Compounds and method for blocking transmission of malarial parasite

ABSTRACT

Disclosed are compounds of formula (I) and formula (II): (I) (II) wherein R 1 , R 2 , A, and B are as defined herein. Also disclosed is a method of blocking transmission of a  Plasmodium  parasite and a method of treating or preventing malaria comprising administering to an animal an effective amount of a first compound of formula (I) or (II) either alone or in combination with a second compound selected from elesclomol, NSC174938, NVP-AUY922, Maduramicin, Narasin, Alvespimycin, Omacetaxine, Thiram, Zinc pyrithione, Phanquinone, Bortezomib, Salinomycin sodium, Monensin sodium, Dipyrithione, Dicyclopentamethylene-thiuram disulfide, YM155, Withaferin A, Adriamycin, Romidepsin, AZD-1152-HQPA, CAY10581, Plicamycin, CUDC-101, Auranofin, Trametinib, GSK-458, Afatinib, and Panobinostat.

CROSS-REFERENCE TO RELATED APPLICATIONS

This patent application is the national stage of International Patent Application No. PCT/US2017/056619, filed Oct. 13, 2017, which claims the benefit of U.S. Provisional Patent Application No. 62/407,861, filed Oct. 13, 2016, the disclosures of which are incorporated by reference in their entirety for all purposes.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with Government support. The Government has certain rights in the invention.

BACKGROUND OF THE INVENTION

Malaria cases and deaths have dropped 50% in 29 countries since 2000 due to the combined effects of long-lasting insecticidal bed nets, indoor residual spraying, and artemisinin-based combination therapies (ACTs) [Kilama W. et al., Lancet, 2009, 374: 1480-1482]. This success has raised hopes for malaria eradication and consequently stimulated interest in developing new reagents that block gametocyte transmission, such as novel and safe gametocytocidal drugs [Buchholz K. et al., The Journal of Infectious Diseases, 2011, 203: 1445-1453]. Previous drug development efforts have focused primarily on the asexual parasites that cause symptoms but not malaria transmission. To be transmitted from person to person via mosquitoes, the parasites must switch from asexual to sexual development and produce male and female gametocytes. Once gametocytes are taken up in a blood meal by a mosquito, fertilization is stimulated and the resulting zygote differentiates into a motile ookinete that migrates across the midgut epithelium of the mosquito and forms an oocyst. Over the course of the next 2 weeks, tens of thousands of infectious sporozoites are generated and sequestered in the mosquito salivary glands until released into a vertebrate host for transmission during the next blood meal.

Sexual stage P. falciparum gametocytes have a lifespan of over 3 weeks and are not cleared effectively by current antimalarial agents, except primaquine (PQ) [Sweeney A W et al., American Journal of Tropical Medicine and Hygiene, 2004, 71: 187-189; Peatey C L et al., Journal of Infectious Diseases, 2009, 200: 1518-1521] which is not widely used because it causes hemolytic anemia in patients with glucose-6-phosphate dehydrogenase deficiency [Baird J K et al., Trends in Parasitology, 2011, 27: 11-16]. Consequently, treatment with current antimalarial drugs often results in asymptomatic carriers who remain infectious for weeks after the clearance of asexual parasites. Despite the risks of PQ, its efficacy with artemisinin combination therapy (ACT) in reducing malaria transmission in the PQ-tolerant patients was recently demonstrated in test regions. Other than PQ, the only other gametocytocidal candidate being tested is methylene blue.

Thus, a new generation of antimalarial agents with potent activities against both sexual and asexual parasites is urgently needed for better therapeutic effect and eradication of malarial infection globally.

BRIEF SUMMARY OF THE INVENTION

The invention provides a compound of formula (I):

wherein A is CR⁵ or N,

B is CR³═CR⁴ or NR⁶,

R¹ is C₆₋₁₀ aryl or heteroaryl substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NHR¹⁵, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, and diazepinyl, or an aryl bicyclic lactam of the formula:

wherein the aryl group of the aryl bicyclic lactam is optionally substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NH₂, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, 2-(dimethylamino)ethyl)amino, dimethylamino, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, azetidinyl and diazepinyl,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-imdazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, N-methyl-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, 4-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-hydroxyphenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 3-cyano-4-chlorophenyl, 3-cyano-5-methylphenyl, 3-hydroxy-4-chlorophenyl, 4-hydroxymethylphenyl, 3-amino-4-chlorophenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, morpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylphenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(diethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, 1-oxoisoindolin-5-yl, 2-amino-4-pyridyl, 3-amino-4-chlorophenyl, 4-aminomethylphenyl, 3-methyl-4-chlorophenyl, 3-methoxy-4-chlorophenyl, 4-aminophenyl, 3-methylphenyl, 3-methoxyphenyl, phenyl, 5-indolinone, 5-isoindolinone, 3-aminophenyl, 2-hydroxy-4-chlorophenyl, 4-cyanophenyl, 4-(2-aminoethyl) phenyl, 4-(2-dimethylaminoethyl) phenyl, 4-azetidinylphenyl, 4-ethylphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-dimethylaminomethylphenyl, 3-cyano-4-fluorophenyl, 3-amino-4-fluorophenyl, 4-(1-hydroxy-1-ethylphenyl, 3-methyl-5-pyridyl, 2-acetylamio-5-pyridyl, 2-oxoindolin-5-yl, benzimidazolin-5-yl, 3-dimethylamino-1-propargyl, 2-pyrrolyl, N-methyl-2-pyrrolyl, 2-thiopheneyl, 3-thiopheneyl, 3-furanyl, 3-aminosulfonylphenyl, 4-dimethylaminomethylphenyl, and 4-pyrrolidinomethylphenyl,

R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl,

R⁵ is hydrogen, C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, hydroxyl, or OR¹⁶,

R⁶ is hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ hydroxyalkyl, C₁₋₆ acyl-C₁₋₆ alkyl, or C₆₋₁₀ aryl,

R⁷-R¹⁰ are independently selected from hydrogen, C₁₋₁₂ alkyl, C₃₋₈ cycloalkyl, CH₂COOR¹³, and H₂N(CH₂)_(n)— wherein n is an integer of 2-6,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing 0 or N atoms, and

R¹³ is C₁₋₁₂ alkyl,

R¹⁵ is hydrogen or C₁₋₁₂ alkyl,

R¹⁶ is hydrogen, C₁₋₁₂ alkyl or C₆₋₁₀ aryl,

or a pharmaceutically acceptable salt thereof.

The invention also provides a compound of formula (II):

wherein A is CR⁵ or N,

wherein R¹ is C₆₋₁₀ aryl or heteroaryl substituted with one or more substituents selected from the group consisting of C₁₋₁₂ alkyl, C₃₋₁₀ cycloalkyl, alkoxy, —CF₃, heterocyclyl, —CONR¹¹R¹², —SO₂NHR¹⁶, and CN,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-amino-5-trifluoromethylpyrimidin-5-yl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-indazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, morpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylphenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, and 1-oxoisoindolin-5-yl,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing O or N atoms,

R¹⁴ is hydrogen or C₁₋₁₂ alkyl, and

R¹⁶ is hydrogen or C₁₋₁₂ alkyl,

or a pharmaceutically acceptable salt thereof.

The invention further provides a pharmaceutical composition comprising a compound or salt of formula (I) or (II) and a pharmaceutically acceptable carrier.

The invention additionally provides a method of blocking transmission of a Plasmodium parasite comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a first compound of formula (I) or (II), optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

The invention also provides a method of treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, the method comprising administering to a mammal a therapeutically effective amount of a first compound of formula (I) or (II), optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

Advantageously, the compounds and methods of the invention compounds kill all stages of malaria parasites.

BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWING(S)

FIG. 1A-1B depict the structures of compounds in accordance with an embodiment of the invention.

FIG. 2AA-2DE depict the structures of additional compounds in accordance with an embodiment of the invention.

DETAILED DESCRIPTION OF THE INVENTION

In an embodiment, the invention provides a compound of formula (I):

wherein A is CR⁵ or N,

B is CR³═CR⁴ or NR⁶,

R¹ is C₆₋₁₀ aryl or heteroaryl substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NHR¹⁵, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, and diazepinyl, or an aryl bicyclic lactam of the formula:

wherein the aryl group of the aryl bicyclic lactam is optionally substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NH₂, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, 2-(dimethylamino)ethyl)amino, dimethylamino, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, azetidinyl and diazepinyl,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-imdazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, N-methyl-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, 4-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-hydroxyphenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 3-cyano-4-chlorophenyl, 3-cyano-5-methylphenyl, 3-hydroxy-4-chlorophenyl, 4-hydroxymethylphenyl, 3-amino-4-chlorophenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, morpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylphenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(diethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, 1-oxoisoindolin-5-yl, 2-amino-4-pyridyl, 3-amino-4-chlorophenyl, 4-aminomethylphenyl, 3-methyl-4-chlorophenyl, 3-methoxy-4-chlorophenyl, 4-aminophenyl, 3-methylphenyl, 3-methoxyphenyl, phenyl, 5-indolinone, 5-isoindolinone, 3-aminophenyl, 2-hydroxy-4-chlorophenyl, 4-cyanophenyl, 4-(2-aminoethyl) phenyl, 4-(2-dimethylaminoethyl) phenyl, 4-azetidinylphenyl, 4-ethylphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-dimethylaminomethylphenyl, 3-cyano-4-fluorophenyl, 3-amino-4-fluorophenyl, 4-(1-hydroxy-1-ethylphenyl, 3-methyl-5-pyridyl, 2-acetylamio-5-pyridyl, 2-oxoindolin-5-yl, benzimidazolin-5-yl, 3-dimethylamino-1-propargyl, 2-pyrrolyl, N-methyl-2-pyrrolyl, 2-thiopheneyl, 3-thiopheneyl, 3-furanyl, 3-aminosulfonylphenyl, 4-dimethylaminomethylphenyl, and 4-pyrrolidinomethylphenyl,

R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl,

R⁵ is hydrogen, C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, hydroxyl, or OR¹⁶,

R⁶ is hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ hydroxyalkyl, C₁₋₆ acyl-C₁₆ alkyl, or C₆₋₁₀ aryl,

R⁷-R¹⁰ are independently selected from hydrogen, C₁₋₁₂ alkyl, C₃₋₈ cycloalkyl, CH₂COOR¹³, and H₂N(CH₂)_(n)— wherein n is an integer of 2-6,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing O or N atoms, and

R¹³ is C₁₋₁₂ alkyl,

R¹⁵ is hydrogen or C₁₋₁₂ alkyl,

R¹⁶ is C₁₋₁₂ alkyl or C₆₋₁₀ aryl

or a pharmaceutically acceptable salt thereof,

with the provisos that when A is CH, B is CR³═CR⁴, R³ and R⁴ are each hydrogen, and R² is 2-amino-5-pyridyl, R¹ is not 3-cyanophenyl or 3-aminosulfonylphenyl.

Referring now to terminology used generically herein, the term “alkyl” means a straight-chain or branched alkyl substituent containing from, for example, 1 to about 6 carbon atoms, preferably from 1 to about 4 carbon atoms, more preferably from 1 to 2 carbon atoms. Examples of such substituents include methyl, ethyl, propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isoamyl, hexyl, and the like.

The term “aryl” refers to an unsubstituted or substituted aromatic carbocyclic substituent, as commonly understood in the art, and the term “C₆-C₁₀ aryl” includes phenyl and naphthyl. It is understood that the term aryl applies to cyclic substituents that are planar and comprise 4n+2 π electrons, according to Hückel's Rule. The aryl group is optionally substituted with 1, 2, 3, 4, or 5 substituents as recited herein such as with alkyl groups such as methyl groups, ethyl groups, and the like, halo, dihaloalkyl, trihaloalkyl, nitro, hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino, acylamino, acylalkylamino, alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, aryloxycarbonyl, cyanomethyl, alkylsulfonyl, aminosulfonyl, alkylaminosulfonyl, dialkylaminosulfonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, thio, alkylthio, arylthio, and the like, wherein the optional substituent can be present at any open position on the aryl group.

The term “heteroaryl” refers to a monocyclic or bicyclic 5 to 10-membered ring system as described herein, wherein the heteroaryl group is unsaturated and satisfies Hückel's rule, and wherein the heteroaryl contains 1-4 heteroatoms independently selected from nitrogen, oxygen and sulfur. Non-limiting examples of suitable heteroaryl groups include furanyl, thiopheneyl, pyrrolyl, pyrazolyl, imidazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, 1,3,4-oxadiazol-2-yl, 1,2,4-oxadiazol-2-yl, 5-methyl-1,3,4-oxadiazole, 3-methyl-1,2,4-oxadiazole, pyridinyl, pyrimidinyl, pyrazinyl, triazinyl, benzofuranyl, benzothiopheneyl, indolyl, indazolyl, imidazolyl, quinolinyl, isoquinolinyl, benzimidazolyl, benzoxazolinyl, benzothiazolinyl, and quinazolinyl. The term “pyridinyl” is synonymous with the term “pyridyl” and both terms refer to an optionally substituted pyridine group. The heteroaryl groups can be attached at any open position on the heteroaryl groups. The terms “heterocyclic” or “heterocyclyl” refer to a 4 to 12-membered heterocyclic ring system as described herein, wherein the heterocycle contains 1 or 2 heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the heterocycle is saturated or monounsaturated. The heterocyclyl or heteroaryl group is optionally substituted with 1, 2, 3, 4, or 5 substituents as recited herein such as with alkyl groups such as methyl groups, ethyl groups, and the like, with hydroxyalkyl groups such as hydroxyl ethyl, or with aryl groups such as phenyl groups, naphthyl groups and the like, wherein the aryl groups can be further substituted with, for example, halo, dihaloalkyl, trihaloalkyl, nitro, hydroxy, alkoxy, aryloxy, amino, alkylamino, dialkylamino, acylamino, acylalkylamino, alkylcarbonyl, alkoxycarbonyl, arylcarbonyl, aryloxycarbonyl, cyanomethyl, alkylsulfonyl, aminosulfonyl, alkylaminosulfonyl, dialkylaminosulfonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, thio, alkylthio, arylthio, and the like, wherein the optional substituent can be present at any open position on the heterocyclyl or heteroaryl group.

The term “acyl” refers to an alkylcarbonyl (R—C(═O)—) substituent. The term “aminosulfonyl” refers to a group of the structure: H₂NSO₂—. The term “alkylsulfonyl” refers to a group of the structure: alkyl-SO₂—. The term “aminocarbonyl” refers to a group of the structure: R¹R²NC(═O)— wherein R¹ and R² are independently hydrogen, alkyl, or aryl.

In any of the above embodiments, A is CR⁵.

In any of the above embodiments, B is CR³═CR⁴.

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —CN.

In certain of these embodiments, R² is selected from 4-chlorophenyl, 4-dimethylaminomethylphenyl, 4-diethylaminomethylphenyl, 2-amino-4-pyridyl, 2-amino-5-pyridyl, and 4-fluorophenyl.

In certain of these embodiments, R¹ is selected from 3-cyanomethylphenyl, 3-cyano-6-methylphenyl, 3-cyano-4-morpholinylphenyl, 3-cyano-2-methylphenyl, 3-cyano-4-piperazinyl-6-methylphenyl, and 3-cyano-4-piperazinylmethylphenyl.

In certain particular embodiments, the compound is selected from:

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —SO₂R¹³.

In certain of these embodiments, R² is selected from 4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 3-amino-4-chlorophenyl, 4-fluorophenyl, 4-dimethylaminomethylphenyl, 4-aminomethylphenyl, 3-methyl-4-chlorophenyl, 3-methoxy-4-chlorophenyl, 3-cyano-4-chlorophenyl, and 4-pyrrolidinomethylphenyl.

In certain of these embodiments, R¹ is selected from 4-methylsulfonylphenyl, 4-ethylsulfonylphenyl, 2-methyl-4-methylsulfonylphenyl, and 3-piperazinylmethyl-4-methylsulfonylphenyl.

In certain particular embodiments, the compound is selected from:

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —SO₂NH₂.

In certain of these embodiments, R² is 4-chlorophenyl or 2-amino-5-pyridiyl.

In certain of these embodiments, R¹ is 4-aminosulfonylphenyl or 3-aminosulfonylphenyl.

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —CONR¹¹R¹².

In certain of these embodiments, R² is selected from 4-chlorophenyl, 4-fluorophenyl, 4-dimethylaminomethylphenyl, 4-aminomethylphenyl, 3-amino-4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 4-aminophenyl, 4-hydroxyphenyl, 4-hydroxymethylphenyl, 3-methylphenyl, 3-methoxyphenyl, 3-cyanophenyl, phenyl, 5-indolinone, 3-hydroxyphenyl, 5-isoindolinone, 3-aminophenyl, 2-hydroxy-4-chlorophenyl, 3-cyano-4-chlorophenyl, 4-cyanophenyl, 4-(2-aminoethyl)phenyl, 4-(2-dimethylaminoethyl)phenyl, 4-azetidinylphenyl, 4-ethylphenyl, 2-amino-5-pyridyl, and 4-(1-aminocycloprop-1-yl)phenyl.

In certain of these embodiments, R¹ is selected from 4-methylcarbonylaminophenyl, 4-morpholinocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-(3,5-dimethylaminomorpholino)carbonylphenyl, 4-(4-methylpiperazinyl)carbonylphenyl, 4-piperazinylcarbonylphenyl, 4-piperidinylcarbonylphenyl, 4-[N,N-bis(2-hydroxyethyl]carbonylphenyl, 4-cyclopentylaminocarbonylphenyl, 4-azetidinylcarbonylphenyl, 4-(4-hydroxyethylpiperazinyl)carbonylphenyl, 2-methyl-4-methylaminocarbonylphenyl, 3-chloro-4-methylaminocarbonylphenyl, 4-cyclopropylaminocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-1-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)carbonylphenyl, 4-1-(3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)carbonylphenyl, 1-(5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)carbonylphenyl, 1-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)carbonylphenyl, 4-(N-cyclopropyl-N-cyclohexylamino)carbonylphenyl, 4-(N-methyl-N-cyclopropylamino)carbonylphenyl, 4-cyclobutylaminocarbonylphenyl, 2trifluoroethylaminocarbonylphenyl, 4-(2-dimethylaminoethylaminocarbonyl)phenyl, 4-(4-[2-dimethylaminoethyl]piperazin-1-yl-carbonyl)phenyl, 4-cyclopropylaminocarbonylphenyl 4-(1-[2-dimethylaminoethyl]piperidin-4-amino)carbonylphenyl, 4-(N-(1-(2-hydroxyethyl)azetidin-3-ylamino)carbonylphenyl, 4-(pyrrolidinyl-3-amino)carbonylphenyl.

In certain particular embodiments, the compound is selected from:

In certain embodiments, R¹ is a aryl bicyclic lactam of the formula:

In certain of these embodiments, R⁷, R⁸, R⁹, and R¹⁰ are hydrogen, C₁₋₁₂ alkyl, or H₂N(CH₂)_(n)— wherein n is an integer of 2-6.

In certain of these embodiments, R² is selected from 4-chlorophenyl, 4-dimethylaminomethylphenyl, 4-diethylaminomethylphenyl, 4-fluorophenyl, 2-amino-5-pyridyl, and 4-(1-aminocycloprop-1-yl)phenyl.

In certain particular embodiments, the compound is selected from:

In any of the above embodiments, B is NR⁶.

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —CN.

In certain of these embodiments, R⁶ is hydrogen or methyl.

In certain of these embodiments, R² is selected from 4-chlorophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-methoxy-4-chlorophenyl, 3-aminophenyl, 3-methylphenyl, 3-dimethylaminomethylphenyl, 3-cyano-4-fluorophenyl, 3-hydroxy-4-fluorophenyl, 3-methoxy-4-fluorophenyl, 3-cyano-4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 4-fluorophenyl, 3-cyanophenyl, 3-hydroxyphenyl, 3-amino-4-fluorophenyl, 4-dimethylaminophenyl, 3-cyanophenyl, 4-methylphenyl, 4-aminophenyl, 4-(1-hydroxy-1-ethyl)phenyl, 4-(2-aminoethylphenyl, 2-amino-5-pyridyl, 2-amino-5-pyrimidyl, 2-methyl-5-pyridyl, 2-acetylamino-5-pyridyl, 2-oxoindolin-5-yl, benzimidazolin-5-yl, 3-dimethylamino-1-propargyl, 4-(2-dimethylaminoethyl)phenyl, 2-pyrrolyl, N-methyl-2-pyrrolyl, 2-thiopheneyl, 3-thiopheneyl, 3-furanyl, 3-aminosulfonylphenyl, and 4-dimethylaminomethylphenyl.

In certain of these embodiments, R¹ is selected from 3-cyano-6-methyl phenyl, 3-cyano-4-piperazinyl-6-methylphenyl, 3-cyanophenyl, 3-cyano-4-piperazinylphenyl, 3-cyano-4-morpholinylphenyl, 4-(4-(2-hydroxyethylpiperazinyl-3-cyanophenyl, 3-cyano-4-(1,4-diazepan-1-yl)phenyl, 3-cyano-4-(4-acetylaminopiperazin-1-yl), 2-methyl-3-cyanophenyl, 3-cyano-4-(3-aminoazetidinyl)-6-methylphenyl, 3-cyano-4-(2-dimethylaminoethyl)-6-methylphenyl, 2-piperazinyl-3-cyano-6-methyl-5-pyridyl, 2-piperazinyl-3-cyano-5-pyridyl, 3-cyano-4-methoxyphenyl, 3-cyano-4-(4-hydroxypiperidinyl)phenyl, 3-cyano-4-(4-aminopiperidinyl)phenyl, and 2-methyl-3-cyanophenyl.

In certain particular embodiments, the compound is selected from:

In certain embodiments, R¹ is C₆₋₁₀ substituted with —SO₂R¹³.

In certain of these embodiments, R² is 2-amino-5-pyridyl or 4-chlorophenyl.

In certain of these embodiments, R¹ is 4-aminosulfonylphenyl.

In certain particular embodiments, the compound is:

In certain embodiments, R¹ is C₆₋₁₀ aryl substituted with —CONR¹¹R¹².

In certain of these embodiments, R² is selected from 4-chlorophenyl, 2-amino-5-pyridyl, 3-hydroxy-4-chlorophenyl, 4-fluorophenyl, 3-cyanophenyl, 4-dimethylaminomethylphenyl, and 4-methylphenyl.

In certain of these embodiments, R¹ is 4-morpholinocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-(N-methyl-N-cyclopropyl)aminocarbonylphenyl, or 4-piperidinocarbonylphenyl.

In certain particular embodiments, the compound is selected from:

In certain embodiments, R¹ is:

In certain of these embodiments, R² is 4-chlorophenyl or 4-dimethylaminophenyl.

In certain preferred embodiments, R¹ is

In an embodiment, the invention provides a compound of formula (II):

wherein A is CR⁵ or N,

B is CR³═CR⁴ or NR⁶,

wherein R¹ is C₆₋₁₀ aryl or heteroaryl substituted with one or more substituents selected from the group consisting of C₁₋₁₂ alkyl, C₃₋₁₀ cycloalkyl, alkoxy, —CF₃, heterocyclyl, —CONR¹¹R¹², —SO₂NHR¹⁶, and CN,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-amino-5-trifluoromethylpyrimidin-5-yl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-indazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, porpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylpenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, and 1-oxoisoindolin-5-yl,

R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl,

R⁵ is hydrogen, C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, hydroxyl, or OR⁵,

R⁶ is hydrogen, C₁₋₁₂ alkyl or C₆₋₁₀ aryl,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing O or N atoms,

R¹⁴ is hydrogen or C₁₋₁₂ alkyl, and

R¹⁶ is hydrogen or C₁₋₁₂ alkyl,

or a pharmaceutically acceptable salt thereof.

In certain preferred embodiments, R¹⁴ is methyl.

In certain of these embodiments, R² is 2-amino-5-pyridyl or 4-chlorophenyl.

In certain of these embodiments, R¹ is selected from 3-trifluoromethylphenyl, 4-methylphenyl, 3-cyanophenyl, 3-pyridyl, 4-pyrrolidinylcarbonylphenyl, 4-cyclopropylaminocarbonylphenyl, 4-(3-dimethylaminopropylaminocarbonyl)phenyl, and 3-trifluoromethyl-4-morpholinocarbonylphenyl.

Chemistry

The compounds of the invention can be synthesized using any suitable route. In an embodiment, the compounds of formula (I) wherein A is CH and B is CR³═CR⁴ can be synthesized by the route shown in Scheme 1.

A substituted 4-amino-6-bromoquinoline-3-carbaldehyde 1 can be reacted with an acetyl chloride in the presence of a base such as diethylaminopropylamine in a solvent such as tetrahydrofuran, optionally under microwave irradiation, to give a 9-bromobenzo[h][1,6]naphthyridin-2(1H)-one 1′. Suzuki coupling of 1′ with an arylboronic acid R¹B(OR″)₂ in the presence of a catalyst such as Pd(Ph₃P)₄ and a base such as NaHCO₃ in a solvent such as DMF/water under microwave irradiation gives the desired compound 2.

In an embodiment, a compound of formula (I) wherein A is CH and B is NR⁶ can be synthesized by the route shown in Scheme 2.

Reagents and conditions. (i) 150° C., 1,4-dioxanes; (ii) SnCl₂, 65° C., 3.5 h; (iii) ClCO₂CCl₃, Et₃N; (iv) MeI, NaH, THF

Reaction of a substituted aniline such as, for example, 3-trifluoromethylaniline with 6-bromo-4-chloro-3-nitroquinoline in a solvent such as 1,4-dioxane gives the substituted quinoline 17. Reduction of the nitro group with a reducing agent such as SnCl₂ gives the amino compound 18. Cyclization of compound 18 with a carbonylation reagent such as triphogene gives the substituted 8-bromo-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one 19. N-alkylation of compound 19 can be performed using an alkylating agent such as methyl iodide in the presence of a base such as NaH in a solvent such as tetrahydrofuran to give the N-alkylated compound 20. Suzuki coupling of compounds 19 and 20 gives 8-functionalized 1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-ones.

The compounds of formula (II) can be prepared by the route shown in Scheme 3.

Reaction of 1-(6-bromo-4-hydroxyquinolin-3-yl)ethan-s 1-one 1 with substituted hydrazine 2 in a solvent such as acetic acid gives substituted 8-bromo-1H-pyrazolo[4,3-c]quinoline 3. Suzuki coupling of 3 with an arylboronic acid 4 in the presence of a catalyst such as Pd(Ph₃P)₄ and a base such as NaHCO₃ in a solvent such as DMF/water under microwave irradiation gives the desired compound 5.

In an embodiment, the invention provides a method for the preparation of heteroaryl-fused imidazolidin-2-ones. In an embodiment, the method comprises the steps of (a) providing a compound of formula 100:

wherein X² is F, Cl, Br, or I, V is CR¹⁰⁴ or N, W is CR¹⁰⁵ or N, Y is CR¹⁰⁶ or N, Z is CR¹⁰⁷ or N, wherein optionally Y and Z, wherein R¹⁰⁴-R¹⁰⁷ are independently hydrogen, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl, or when Y is CR¹⁰⁶ and Z is CR¹⁰⁷, R¹⁰⁶ and R¹⁰⁶, taken together with the carbons to which they are bound, form a fused 5- to 8-membered carbocyclic, aryl, heterocyclyl, or heteroaryl ring, wherein the heterocyclyl contains one or more atoms selected from N, O, and S, wherein at least one of V, W, Y, and Z is N, wherein R¹⁰⁰ is alkyl or aryl, wherein R¹⁰¹ is hydrogen, alkyl, or aryl, and (b) reacting the compound of formula 100 with an amine of the formula: R¹⁰²NH₂ to give a compound of the formula 101:

wherein R¹⁰² is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted heterocyclylalkyl, optionally substituted cycloalkylalkyl, or optionally substituted heteroarylalkyl.

The method comprises mixing ortho-halo heteroaryl carbamates and a primary amine under conditions such that an imidazolidin-2-one ring is formed. In typical embodiments, an ortho-halo heteroaryl carbamate 100 reacts with a primary amine to give a heteroaryl-fused imidazolidin-2-one 101 as the product (Scheme 4). The reaction usually takes place in preferred acidic conditions under conventional heat or microwave heat. When a polycyclic ortho-halo heteroaryl carbamate 100′ is used as the starting material, a polycyclic imidazolidin-2-one product 101′ is formed (Scheme 5). The acid used here can be an organic or inorganic acid such as hydrochloric acid (HCl), nitric acid (HNO₃), phosphoric acid (H₃PO₄), sulphuric acid (H₂SO₄), boric acid (H₃BO₃), hydrofluoric acid (HF), hydrobromic acid (HBr), perchloric acid (HClO₄), formic acid (HCOOH), acetic acid (CH₃COOH), propanoic acid (CH₃CH₂COOH), and trifluoroacetic acid (TFA). The solvent used here can be a high boiling point solvent such as 2-pentanol, 3-pentanol, cyclohexanol, polyethylene glycol (PEG), 2-methyl-2-butanol, tert-butanol, MeCN, 1,2-dimethoxyethane (DME), 1,2-dichloroethane (DCE), 1,4-dioxane, toluene, N,N-dimethylacetamide (DMA), N,N-dimethylformamide (DMF), 1-methyl-2-pyrrolidinone (NMP), xylenes, dimethylsulfoxide (DMSO), o-dichlorobenezene (o-DCB) and ionic liquid. The reaction temperature is between 25° C. to 250° C.

In an embodiment, the method is useful for the synthesis of compounds of formula (I) wherein B is NR⁶.

The synthesis of carbamate 104 and N-alkyl carbamate 105 can be accomplished as shown in Scheme 6. Reduction of nitro quinoline 102 wherein X¹ and X² are independently halo or OSO₂R wherein R is alkyl, CF₃, or optionally substituted aryl with a suitable reducing agent such as SnCl₂ or under catalytic hydrogenation conditions provides amino quinoline 103. Reaction of amino quinolone 103 with an alkyl or aryl chloroformate provides carbamate 104. Carbamate 104 can be alkylated, for example, using an alkyl halide or alkyl sulfonate in the presence of a base such as sodium hydride provides N-alkyl carbamate 105.

Reagents and conditions: (a) SnCl₂*2H₂O, AcOH, EtOH, rfx, 3 h, 64%; (b) ClCOOiPr, pyr, DCM, rt, 4 h, 66%; (c) MeI, NaH, THF, rt, 18 h, 93%.

Reaction of N-alkyl carbamate 105 with an optionally substituted arylamine or alkylamine in a solvent such as 3-pentanol under microwave irradiation provides 1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one 106 as shown in Scheme 7. 1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one 106 reacts with aryl boronic acids R¹⁰³B(OH)₂ under Suzuki coupling conditions in the presence of a catalyst such as Pd(PPh₃)₄ and a base such as K₃PO₄ in a solvent or solvent mixture such as DMF/H₂O/i-PrOH under microwave irradiation to give 8-arylated 1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one 107.

Heteroaryl-fused imidazolidin-2-ones are important motifs in the biologically active compounds. For example, the dual PI3K/mTOR inhibitors, NVP-BEZ235 and NVP-BGT226, have both been studied in clinic trials for their potential as future anti-cancer agents. The synthesis of heteroaryl-fused imidazolidin-2-ones usually involves the reaction of phosgene or its derivatives with heteroaryl 1,2-diamines to form the cyclic urea rings. Alternatively, heteroaryl-fused imidazolidin-2-ones were prepared from their corresponding ortho-chloro heteroaryl amines via Pd-catalyzed Buchwald-Hartwig C—N bond formation followed by an intramolecular cyclization. These methods use either toxic phosgene derivatives or expensive transition-metal catalysts. A general, metal-free method using non-toxic reagents is desired for the synthesis of biologically interesting molecules containing polycyclic imidazolidin-2-ones.

NVP-BEZ235 and NVP-BGT226 have both been studied in clinical trials for their potential as future anti-cancer agents. The N-methyl i-Pr carbamate 110 could react satisfactorily with two equivalents of aniline 111 and three equivalents of HCl, subsequent one-pot coupling gave desired 112 (NVP-BEZ235) in 85% yield (Scheme 8). When Boc-protected CF₃-substituted aniline 113 was used under optimal condition, the desired product 114 (NVP-BGT226) was obtained as a free base in 42% yield. Additionally, such transformation has been effected on a gram scale under conventional heating. N-methyl i-Pr carbamate 110 reacts with two equivalents of Boc-protected CF₃-substituted aniline 113 and ten equivalents of HCl in high boiling point solvent such as N-Methyl-pyrrolidone (NMP), subsequent one-pot coupling furnished desired 114 (NVP-BGT226) in 77% yield (Scheme 8).

The described new method for the synthesis of heteroaryl-fused imidazolidin-2-ones can be applied to the construction of other pharmaceutically active molecules such as I-BET151 (GSK1210151A), a novel small molecule inhibitor of the BET family, which has been investigated substantially in preclinical models for the treatment of mixed lineage leukemia (MLL). Telcagepant, also known as MK0974, is an orally bioavailable calcitonin gene-related peptide (CGRP) receptor antagonist under development for the acute treatment and prevention of migraine (Scheme 9).

In accordance with an embodiment of the invention, any of the compounds or salts thereof can be administered in the form of a pharmaceutical composition comprising the compound and a pharmaceutically acceptable carrier.

The pharmaceutically acceptable carriers described herein, for example, vehicles, adjuvants, excipients, or diluents, are well known to those who are skilled in the art and are readily available to the public. It is preferred that the pharmaceutically acceptable carrier be one which is chemically inert to the active compounds and one which has no detrimental side effects or toxicity under the conditions of use.

The choice of carrier will be determined in part by the particular active agent, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of the pharmaceutical composition of the present invention. The following formulations for oral, aerosol, parenteral, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, intrathecal, rectal, and vaginal administration are merely exemplary and are in no way limiting.

Formulations suitable for oral administration can consist of (a) liquid solutions, such as an effective amount of the compound dissolved in diluents, such as water, saline, or orange juice; (b) capsules, sachets, tablets, lozenges, and troches, each containing a predetermined amount of the active ingredient, as solids or granules; (c) powders; (d) suspensions in an appropriate liquid; and (e) suitable emulsions. Liquid formulations may include diluents, such as water and alcohols, for example, ethanol, benzyl alcohol, and the polyethylene alcohols, either with or without the addition of a pharmaceutically acceptable surfactant, suspending agent, or emulsifying agent. Capsule forms can be of the ordinary hard- or soft-shelled gelatin type containing, for example, surfactants, lubricants, and inert fillers, such as lactose, sucrose, calcium phosphate, and cornstarch. Tablet forms can include one or more of lactose, sucrose, mannitol, corn starch, potato starch, alginic acid, microcrystalline cellulose, acacia, gelatin, guar gum, colloidal silicon dioxide, croscarmellose sodium, talc, magnesium stearate, calcium stearate, zinc stearate, stearic acid, and other excipients, colorants, diluents, buffering agents, disintegrating agents, moistening agents, preservatives, flavoring agents, and pharmacologically compatible carriers. Lozenge forms can comprise the active ingredient in a flavor, usually sucrose and acacia or tragacanth, as well as pastilles comprising the active ingredient in an inert base, such as gelatin and glycerin, or sucrose and acacia, emulsions, gels, and the like containing, in addition to the active ingredient, such carriers as are known in the art.

The compounds of the present invention, alone or in combination with other suitable components, can be made into aerosol formulations to be administered via inhalation. These aerosol formulations can be placed into pressurized acceptable propellants, such as dichlorodifluoromethane, propane, nitrogen, and the like. They also may be formulated as pharmaceuticals for non-pressured preparations, such as in a nebulizer or an atomizer.

Formulations suitable for parenteral administration include aqueous and non-aqueous, isotonic sterile injection solutions, which can contain anti-oxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions that can include suspending agents, solubilizers, thickening agents, stabilizers, and preservatives. The compound can be administered in a physiologically acceptable diluent in a pharmaceutical carrier, such as a sterile liquid or mixture of liquids, including water, saline, aqueous dextrose and related sugar solutions, an alcohol, such as ethanol, isopropanol, or hexadecyl alcohol, glycols, such as propylene glycol or polyethylene glycol, glycerol ketals, such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers, such as poly(ethyleneglycol) 400, an oil, a fatty acid, a fatty acid ester or glyceride, or an acetylated fatty acid glyceride with or without the addition of a pharmaceutically acceptable surfactant, such as a soap or a detergent, suspending agent, such as pectin, carbomers, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose, or emulsifying agents and other pharmaceutical adjuvants.

Oils, which can be used in parenteral formulations include petroleum, animal, vegetable, or synthetic oils. Specific examples of oils include peanut, soybean, sesame, cottonseed, corn, olive, petrolatum, and mineral. Suitable fatty acids for use in parenteral formulations include oleic acid, stearic acid, and isostearic acid. Ethyl oleate and isopropyl myristate are examples of suitable fatty acid esters. Suitable soaps for use in parenteral formulations include fatty alkali metal, ammonium, and triethanolamine salts, and suitable detergents include (a) cationic detergents such as, for example, dimethyl dialkyl ammonium halides, and alkyl pyridinium halides, (b) anionic detergents such as, for example, alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates, and sulfosuccinates, (c) nonionic detergents such as, for example, fatty amine oxides, fatty acid alkanolamides, and polyoxyethylene-polypropylene copolymers, (d) amphoteric detergents such as, for example, alkyl-beta-aminopropionates, and 2-alkyl-imidazoline quaternary ammonium salts, and (3) mixtures thereof.

The parenteral formulations typically contain from about 0.5 to about 25% by weight of the active ingredient in solution. Suitable preservatives and buffers can be used in such formulations. In order to minimize or eliminate irritation at the site of injection, such compositions may contain one or more nonionic surfactants having a hydrophile-lipophile balance (HLB) of from about 12 to about 17. The quantity of surfactant in such formulations ranges from about 5 to about 15% by weight. Suitable surfactants include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate and the high molecular weight adducts of ethylene oxide with a hydrophobic base, formed by the condensation of propylene oxide with propylene glycol. The parenteral formulations can be presented in unit-dose or multi-dose sealed containers, such as ampoules and vials, and can be stored in a freeze-dried (lyophilized) condition requiring only the addition of the sterile liquid carrier, for example, water, for injections, immediately prior to use. Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules, and tablets of the kind previously described.

The compounds of the present invention may be made into injectable formulations. The requirements for effective pharmaceutical carriers for injectable compositions are well known to those of ordinary skill in the art. See Pharmaceutics and Pharmacy Practice, J. B. Lippincott Co., Philadelphia, Pa., Banker and Chalmers, eds., pages 238-250 (1982), and ASHP Handbook on Injectable Drugs, Toissel, 4th ed., pages 622-630 (1986).

Additionally, the compounds of the present invention may be made into suppositories by mixing with a variety of bases, such as emulsifying bases or water-soluble bases. Formulations suitable for vaginal administration may be presented as pessaries, tampons, creams, gels, pastes, foams, or spray formulas containing, in addition to the active ingredient, such carriers as are known in the art to be appropriate.

Suitable carriers and their formulations are further described in A. R. Gennaro, ed., Remington: The Science and Practice of Pharmacy (19th ed.), Mack Publishing Company, Easton, Pa. (1995).

The compound of the invention or a composition thereof can be administered as a pharmaceutically acceptable acid-addition, base neutralized or addition salt, formed by reaction with inorganic acids, such as hydrochloric acid, hydrobromic acid, perchloric acid, nitric acid, thiocyanic acid, sulfuric acid, and phosphoric acid, and organic acids such as formic acid, acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid, malonic acid, succinic acid, maleic acid, and fumaric acid, or by reaction with an inorganic base, such as sodium hydroxide, ammonium hydroxide, potassium hydroxide, and organic bases, such as mono-, di-, trialkyl, and aryl amines and substituted ethanolamines. The conversion to a salt is accomplished by treatment of the base compound with at least a stoichiometric amount of an appropriate acid. Typically, the free base is dissolved in an inert organic solvent such as diethyl ether, ethyl acetate, chloroform, ethanol, methanol, and the like, and the acid is added in a similar solvent. The mixture is maintained at a suitable temperature (e.g., between 0° C. and 50° C.). The resulting salt precipitates spontaneously or can be brought out of solution with a less polar solvent.

The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the parent compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.

It should be recognized that the particular counterion forming a part of any salt of this invention is usually not of a critical nature, so long as the salt as a whole is pharmacologically acceptable and as long as the counterion does not contribute undesired qualities to the salt as a whole.

It is further understood that the above compounds and salts may form solvates, or exist in a substantially uncomplexed form, such as the anhydrous form. As used herein, the term “solvate” refers to a molecular complex wherein the solvent molecule, such as the crystallizing solvent, is incorporated into the crystal lattice. When the solvent incorporated in the solvate is water, the molecular complex is called a hydrate. Pharmaceutically acceptable solvates include hydrates, alcoholates such as methanolates and ethanolates, acetonitrilates and the like. These compounds can also exist in polymorphic forms.

In an embodiment, the invention provides a method of blocking transmission of a Plasmodium parasite comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a compound of the invention. In another embodiment, the invention provides a method of treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, the method comprising administering to a mammal a therapeutically effective amount of a compound of the invention.

The Plasmodium parasite can be any suitable Plasmodium parasite. Non-limiting examples of suitable Plasmodium parasites include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale, and Plasmodium knowlesi. In a preferred embodiment, the Plasmodium parasite is Plasmodium falciparum.

In an embodiment, the Plasmodium parasite is a Plasmodium gametocyte.

In embodiments, the Plasmodium gametocyte is a mature stage II-V gametocyte. In a preferred embodiment, the Plasmodium gametocyte is a stage III-V gametocyte, e.g., a mature stage III-V gametocyte. In another preferred embodiment, the Plasmodium gametocyte is a mature stage V gametocyte.

In certain preferred embodiments, the compound effectively kills Plasmodium gametocytes.

In embodiments, the Plasmodium parasite is a drug-resistant strain. Examples of drug-resistant strains of Plasmodium are described in Kun, J. F. J. et al., Antimicrob Agents Chemother., 1999 September; 43(9): 2205-2208, and references cited therein.

In embodiments, the Plasmodium parasite is in an asexual stage. For example, the Plasmodium parasite can be a sporozoite, a liver stage parasite, a merozoite, an asexual erythrocyte-stage parasite, a zygote, an ookinete, or an oocyst.

The amount or dose of a compound of the invention or a salt thereof, or a composition thereof should be sufficient to effect a therapeutic or prophylactic response in the mammal. The appropriate dose will depend upon several factors. For instance, the dose also will be determined by the existence, nature and extent of any adverse side effects that might accompany the administration of a particular compound or salt. Ultimately, the attending physician will decide the dosage of the compound of the present invention with which to treat each individual patient, taking into consideration a variety of factors, such as age, body weight, general health, diet, sex, compound or salt to be administered, route of administration, and the severity of the condition being treated. By way of example and not intending to limit the invention, the dose of the compound(s) described herein can be about 0.1 mg to about 1 g daily, for example, about 5 mg to about 500 mg daily. Further examples of doses include but are not limited to: 0.1 mg, 0.15 mg, 0.2 mg, 0.25 mg, 0.5 mg, 0.6 mg, 0.75 mg, 1 mg, 1.5 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 12 mg, 15 mg, 17 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 60 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 125 mg, 140 mg, 150 mg, 175 mg, 200 mg, 225 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 950 mg, or 1000 mg/kg body weight per day.

In certain embodiments, the method further comprises administering to the mammal at least one additional antimalarial compound. Any suitable antimalarial compound can be used, many of which are well known in the art. Non-limiting examples of suitable antimalarial compounds include primaquine, bulaquine, artemisinin and derivatives thereof, chloroquine, mefloquine, amodiaquine, piperaquine, pyronaridine, atovaquone, tafenoquine, methylene blue, trioxaquines, endoperoxides such as OZ 439 and OZ 277, decoquinate, 9-anilinoacridines, HIV-protease inhibitors, and natural products such as neem, epoxomicin, harmonine, and riboflavin. In certain preferred embodiments, the compound of the invention is administered in combination with elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, panobinostat, or any combination thereof.

ILLUSTRATIVE EXAMPLES OF EMBODIMENTS

The invention contains the following embodiments:

1. A compound of formula (I):

wherein A is CR⁵ or N,

B is CR³═CR⁴ or NR⁶,

R¹ is C₆₋₁₀ aryl or heteroaryl substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NHR¹⁵, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, and diazepinyl, or an aryl bicyclic lactam of the formula:

wherein the aryl group of the aryl bicyclic lactam is optionally substituted with at least one substituent selected from —CN, cyanomethyl, —SO₂R¹³, —SO₂NH₂, and —CONR¹¹R¹², optionally further in combination with one or more substituents selected from halo, C₁₋₁₂ alkyl, alkoxy, 2-(dimethylamino)ethyl)amino, dimethylamino, a heterocyclyl group selected from the group consisting of optionally substituted piperazinyl, morpholinyl, pyrrolinidyl, azetidinyl and diazepinyl,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-imdazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, N-methyl-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, 4-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-hydroxyphenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-fluorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 3-cyano-4-chlorophenyl, 3-cyano-5-methylphenyl, 3-hydroxy-4-chlorophenyl, 4-hydroxymethylphenyl, 3-amino-4-chlorophenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, morpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylphenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(diethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, 1-oxoisoindolin-5-yl, 2-amino-4-pyridyl, 3-amino-4-chlorophenyl, 4-aminomethylphenyl, 3-methyl-4-chlorophenyl, 3-methoxy-4-chlorophenyl, 4-aminophenyl, 3-methylphenyl, 3-methoxyphenyl, phenyl, 5-indolinone, 5-isoindolinone, 3-aminophenyl, 2-hydroxy-4-chlorophenyl, 4-cyanophenyl, 4-(2-aminoethyl) phenyl, 4-(2-dimethylaminoethyl) phenyl, 4-azetidinylphenyl, 4-ethylphenyl, 3-fluorophenyl, 3-chlorophenyl, 3-dimethylaminomethylphenyl, 3-cyano-4-fluorophenyl, 3-amino-4-fluorophenyl, 4-(1-hydroxy-1-ethylphenyl, 3-methyl-5-pyridyl, 2-acetylamio-5-pyridyl, 2-oxoindolin-5-yl, benzimidazolin-5-yl, 3-dimethylamino-1-propargyl, 2-pyrrolyl, N-methyl-2-pyrrolyl, 2-thiopheneyl, 3-thiopheneyl, 3-furanyl, 3-aminosulfonylphenyl, 4-dimethylaminomethylphenyl, and 4-pyrrolidinomethylphenyl,

R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl,

R⁵ is hydrogen, C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, hydroxyl, or OR¹⁶,

R⁶ is hydrogen, C₁₋₁₂ alkyl, C₂₋₁₂ hydroxyalkyl, C₁₋₆ acyl-C₁₋₆ alkyl, or C₆₋₁₀ aryl, R⁷-R¹⁰ are independently selected from hydrogen, C₁₋₁₂ alkyl, C₃₋₈ cycloalkyl, CH₂COOR¹³, and H₂N(CH₂)_(n)— wherein n is an integer of 2-6,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing O or N atoms, and

R¹³ is C₁₋₁₂ alkyl,

R¹⁵ is hydrogen or C₁₋₁₂ alkyl,

R¹⁶ is C₁₋₁₂ alkyl, C₁₋₆ acyl, or C₆₋₁₀ aryl,

or a pharmaceutically acceptable salt thereof, with the provisos that when A is CH, B is CR³═CR⁴, R³ and R⁴ are each hydrogen, and R² is 2-amino-5-pyridyl, R¹ is not 3-cyanophenyl or 3-aminosulfonylphenyl.

2. The compound or salt of embodiment 1, wherein A is CR⁵.

3. The compound or salt of embodiment 2, wherein B is CR³═CR⁴.

4. The compound or salt of embodiment 3, wherein R¹ is C₆₋₁₀ aryl substituted with —CN.

5. The compound or salt of embodiment 4, wherein R² is selected from 4-chlorophenyl, 4-dimethylaminomethylphenyl, 4-diethylaminomethylphenyl, 2-amino-4-pyridyl, 2-amino-5-pyridyl, and 4-fluorophenyl.

6. The compound or salt of embodiment 4 or 5, wherein R¹ is selected from 3-cyanomethylphenyl, 3-cyano-6-methylphenyl, 3-cyano-4-morpholinylphenyl, 3-cyano-2-methylphenyl, 3-cyano-4-piperazinyl-6-methylphenyl, and 3-cyano-4-piperazinylmethylphenyl.

7. The compound or salt of any one of embodiments 3-6, wherein the compound is selected from:

8. The compound or salt of embodiment 3, wherein R¹ is C₆₋₁₀ aryl substituted with —SO₂R¹³.

9. The compound or salt of embodiment 8, wherein R² is selected from 4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 3-amino-4-chlorophenyl, 4-fluorophenyl, 4-dimethylaminomethylphenyl, 4-aminomethylphenyl, 3-methyl-4-chlorophenyl, 3-methoxy-4-chlorophenyl, 3-cyano-4-chlorophenyl, and 4-pyrrolidinomethylphenyl.

10. The compound or salt of embodiment 8 or 9, wherein R¹ is selected from 4-methylsulfonylphenyl, 4-ethylsulfonylphenyl, 2-methyl-4-methylsulfonylphenyl, and 3-piperazinylmethyl-4-methylsulfonylphenyl.

11. The compound or salt of any one of embodiments 8-10, wherein the compound is selected from:

12. The compound or salt of embodiment 3, wherein R¹ is C₆₋₁₀ aryl substituted with —SO₂NH₂.

13. The compound or salt of embodiment 12, wherein R² is 4-chlorophenyl or 2-amino-5-pyridiyl.

14. The compound or salt of embodiment 12 or 13, wherein R is 4-aminomethylphenyl or 3-aminomethylphenyl.

15. The compound or salt of embodiment 3, wherein R¹ is C₆₋₁₀ aryl substituted with —CONR¹¹R¹².

16. The compound or salt of embodiment 15, wherein R² is selected from 4-chlorophenyl, 4-fluorophenyl, 4-dimethylaminomethylphenyl, 4-aminomethylphenyl, 3-amino-4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 4-aminophenyl, 4-hydroxyphenyl, 4-hydroxymethylphenyl, 3-methylphenyl, 3-methoxyphenyl, 3-cyanophenyl, phenyl, 5-indolinone, 3-hydroxyphenyl, 5-isoindolinone, 3-aminophenyl, 2-hydroxy-4-chlorophenyl, 3-cyano-4-chlorophenyl, 4-cyanophenyl, 4-(2-aminoethyl)phenyl, 4-(2-dimethylaminoethyl)phenyl, 4-azetidinylphenyl, 4-ethylphenyl, 2-amino-5-pyridyl, and 4-(1-aminocycloprop-1-yl)phenyl.

17. The compound or salt of embodiment 15 or 16, wherein R¹ is selected from 4-methylcarbonylaminophenyl, 4-morpholinocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-(3,5-dimethylaminomorpholino)carbonylphenyl, 4-(4-methylpiperazinyl)carbonylphenyl, 4-piperazinylcarbonylphenyl, 4-piperidinylcarbonylphenyl, 4-[N,N-bis(2-hydroxyethyl]carbonylphenyl, 4-cyclopentylaminocarbonylphenyl, 4-azetidinylcarbonylphenyl, 4-(4-hydroxyethylpiperazinyl)carbonylphenyl, 2-methyl-4-methylaminocarbonylphenyl, 3-chloro-4-methylaminocarbonylphenyl, 4-cyclopropylaminocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-1-(8-oxa-3-azabicyclo[3.2.1]octan-3-yl)carbonylphenyl, 4-1-(3-hydroxy-8-azabicyclo[3.2.1]octan-8-yl)carbonylphenyl, 1-(5-methyl-2,5-diazabicyclo[2.2.1]heptan-2-yl)carbonylphenyl, 1-(5-methylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)carbonylphenyl, 4-(N-cyclopropyl-N-cyclohexylamino)carbonylphenyl, 4-(N-methyl-N-cyclopropylamino)carbonylphenyl, 4-cyclobutylaminocarbonylphenyl, 2trifluoroethylaminocarbonylphenyl, 4-(2-dimethylaminoethylaminocarbonyl)phenyl, 4-(4-[2-dimethylaminoethyl]piperazin-1-yl-carbonyl)phenyl, 4-cyclopropylaminocarbonylphenyl 4-(1-[2-dimethylaminoethyl]piperidin-4-amino)carbonylphenyl, 4-(N-(1-(2-hydroxyethyl)azetidin-3-ylamino)carbonylphenyl, 4-(pyrrolidinyl-3-amino)carbonylphenyl.

18. The compound or salt of any one of embodiments 15-17, wherein the compound is selected from:

19. The compound or salt of embodiment 3, wherein R¹ is a aryl bicyclic lactam of the formula:

20. The compound or salt of embodiment 19, wherein R⁷, R⁸, R⁹, and R¹⁰ are hydrogen, C₁₋₁₂ alkyl, or H₂N(CH₂)_(n)— wherein n is an integer of 2-6.

21. The compound or salt of embodiment 19 or 20, wherein R² is selected from 4-chlorophenyl, 4-dimethylaminomethylphenyl, 4-diethylaminomethylphenyl, 4-fluorophenyl, 2-amino-5-pyridyl, and 4-(1-aminocycloprop-1-yl)phenyl.

22. The compound or salt of any one of embodiments 19-21, wherein the compound is selected from:

23. The compound or salt of embodiment 3, wherein B is NR⁶.

24. The compound or salt of embodiment 23, wherein R¹ is C₆₋₁₀ aryl substituted with —CN.

25. The compound or salt of embodiment 24, wherein R⁶ is hydrogen or methyl.

26. The compound or salt of embodiment 24 or 25, wherein R² is selected from 4-chlorophenyl, 3-fluorophenyl, 3-chlorophenyl, 3-methoxy-4-chlorophenyl, 3-aminophenyl, 3-methylphenyl, 3-dimethylaminomethylphenyl, 3-cyano-4-fluorophenyl, 3-hydroxy-4-fluorophenyl, 3-methoxy-4-fluorophenyl, 3-cyano-4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 4-fluorophenyl, 3-cyanophenyl, 3-hydroxyphenyl, 3-amino-4-fluorophenyl, 4-dimethylaminophenyl, 3-cyanophenyl, 4-methylphenyl, 4-aminophenyl, 4-(1-hydroxy-1-ethyl)phenyl, 4-(2-aminoethyl)phenyl, 2-amino-5-pyridyl, 2-amino-5-pyrimidyl, 2-methyl-5-pyridyl, 2-acetylamino-5-pyridyl, 2-oxoindolin-5-yl, benzimidazolin-5-yl, 3-dimethylamino-1-propargyl, 4-(2-dimethylaminoethyl)phenyl, 2-pyrrolyl, N-methyl-2-pyrrolyl, 2-thiopheneyl, 3-thiopheneyl, 3-furanyl, 3-aminosulfonylphenyl, and 4-dimethylaminomethylphenyl.

27. The compound or salt of any one of embodiments 24-26, wherein R¹ is selected from 3-cyano-6-methyl phenyl, 3-cyano-4-piperazinyl-6-methylphenyl, 3-cyanophenyl, 3-cyano-4-piperazinylphenyl, 3-cyano-4-morpholinylphenyl, 4-(4-(2-hydroxyethylpiperazinyl-3-cyanophenyl, 3-cyano-4-(1,4-diazepan-1-yl)phenyl, 3-cyano-4-(4-acetylaminopiperazin-1-yl), 2-methyl-3-cyanophenyl, 3-cyano-4-(3-aminoazetidinyl)-6-methylphenyl, 3-cyano-4-(2-dimethylaminoethyl)-6-methylphenyl, 2-piperazinyl-3-cyano-6-methyl-5-pyridyl, 2-piperazinyl-3-cyano-5-pyridyl, 3-cyano-4-methoxyphenyl, 3-cyano-4-(4-hydroxypiperidinyl)phenyl, 3-cyano-4-(4-aminopiperidinyl)phenyl, and 2-methyl-3-cyanophenyl.

28. The compound or salt of any one of embodiments 24-27, wherein the compound is selected from:

29. The compound or salt of embodiment 23, wherein R¹ is C₆₋₁₀ aryl substituted with —S₂R¹³.

30. The compound or salt of embodiment 29, wherein R² is 2-amino-5-pyridyl or 4-chlorophenyl.

31. The compound or salt of embodiment 29 or 30 wherein R¹ is 4-aminosulfonylphenyl.

32. The compound or salt of any one of embodiments 29-31, wherein the compound is:

33. The compound or salt of embodiment 23, wherein R¹ is C₆₋₁₀ aryl substituted with —CONR¹¹R¹².

34. The compound or salt of embodiment 33, wherein R² is selected from 4-chlorophenyl, 2-amino-5-pyridyl, 3-hydroxy-4-chlorophenyl, 4-fluorophenyl, 3-cyanophenyl, 4-dimethylaminomethylphenyl, and 4-methylphenyl.

35. The compound or salt of embodiment 33 or 34, wherein R¹ is 4-morpholinocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-(N-methyl-N-cyclopropyl)aminocarbonylphenyl, or 4-piperidinocarbonylphenyl.

36. The compound or salt of any one of embodiments 33-35, wherein the compound is selected from:

37. The compound or salt of embodiment 23, wherein R¹ is:

38. The compound or salt of embodiment 37, wherein R² is 4-chlorophenyl or 4-dimethylaminophenyl.

39. The compound or salt of embodiment 38, wherein R¹ is

40. A pharmaceutical composition comprising a compound or salt of any one of embodiments 1-39 and a pharmaceutically acceptable carrier.

41. A method of blocking transmission of a Plasmodium parasite comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a first compound of any one of embodiments 1-39, optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

42. A method of treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, the method comprising administering to a mammal a therapeutically effective amount of a first compound of any one of embodiments 1-39, optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

43. A compound of any one of embodiments 1-39 for use in blocking transmission of a Plasmodium parasite in a mammal in need of therefore, optionally for use in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

44. A compound of any one of embodiments 1-39 for use in killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, optionally for use in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

45. A compound of formula (II):

wherein A is CR⁵ or N,

wherein R¹ is C₆₋₁₀ aryl or heteroaryl substituted with one or more substituents selected from the group consisting of C₁₋₁₂ alkyl, C₃₋₁₀ cycloalkyl, alkoxy, —CF₃, heterocyclyl, —CONR¹¹R¹², —SO₂NHR¹⁶, and CN,

R² is selected from 2-amino-5-pyridinyl, 4-pyridinyl, 2-amino-5-pyrimidinyl, 3-pyridyl, quinolin-3-yl, 5-pyrimidinyl, 2-acetylamino-5-pyridyl, 2-amino-4-methylpyrimidin-5-yl, 1-piperazinyl, indol-5-yl, 1H-indazol-5-yl, 4-aminophenyl, 1,2,3,6-tetrahydropyridin-4-yl, 1H-pyrazol-4-yl, 1H-benzo[d]imidazol-5-yl, 4-sulfonylaminophenyl, 2-dimethylaminopyrimidin-5-yl, 3-trifluoromethylphenyl, bromo, 3-aminophenyl, vinyl, 4-aminocarbonylphenyl, 3-cyanophenyl, 3-trifluoromethyl-5-pyridyl, tetrazolyl, 4-chlorophenyl, 4-methoxyphenyl, 3-aminocarbonylphenyl, 3-acetylphenyl, 2,3-dihydrobenzofuran-6-yl, 1-methyl-1H-indol-5-yl, benzo[d][1,3]dioxo-5-yl, 4-fluorophenyl, 4-hydroxyphenyl, morpholin-1-yl, benzo[b]thiophen-1-yl, 4-methylsulfonylphenyl, benzo[c][1,2,5]oxadiazol-5-yl, 2-(piperidin-1-yl)-3-pyridinyl, 4-carboxyphenyl, 2-methyl-5-pyridyl, 4-methylsulfonylphenyl, 4-dimethylaminocarbonylphenyl, 4-phenylphenyl, 4-methylphenyl, 3-chloro-5-pyridyl, (3-pyrrolidin-1-yl)phenyl, 4-([piperizin-1-yl]carbonyl)phenyl, 4-([morpholin-1-yl]carbonyl)phenyl, 2-hydroxypyrimidin-5-yl, 3-aminosulfonylphenyl, 2-oxo-1,2,3,4,tetrahydroisoquinolin-6-yl, 2-oxo-1,2,3,4,-tetrahydroquinolin-6-yl, 4-(aminomethyl)phenyl, 4-(dimethylaminomethyl)phenyl, 4-(methylaminocarbonyl)phenyl, 1-oxoindolin-5-yl, 2-oxoindolin-5-yl, and 1-oxoisoindolin-5-yl,

R¹¹ and R¹² selected from the group consisting of hydrogen, C₁₋₁₂ alkyl and C₃₋₁₀ cycloalkyl or, taken together with the N to which they are bound, form an optionally substituted 4-7 membered heterocyclyl ring containing O or N atoms,

R¹⁴ is hydrogen or C₁₋₁₂ alkyl, and

R¹⁶ is hydrogen or C₁₋₁₂ alkyl,

or a pharmaceutically acceptable salt thereof.

46. The compound or salt of embodiment 45, wherein R¹⁴ is methyl.

47. The compound or salt of embodiment 45 or 46, wherein R² is 2-amino-5-pyridyl or 4-chlorophenyl.

48. The compound or salt of any one of embodiments 45-47, wherein R¹ is selected from 3-trifluoromethylphenyl, 4-methylphenyl, 3-cyanophenyl, 3-pyridyl, 4-pyrrolidinylcarbonylphenyl, 4-cyclopropylaminocarbonylphenyl, 4-(3-dimethylaminopropylaminocarbonyl)phenyl, and 3-trifluoromethyl-4-morpholinocarbonylphenyl.

49. A pharmaceutical composition comprising a compound or salt of any one of embodiments 45-48 and a pharmaceutically acceptable carrier.

50. A method of blocking transmission of a Plasmodium parasite comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a first compound of any one of embodiments 45-48, optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

51. A method of treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, the method comprising administering to a mammal a therapeutically effective amount of a first compound of any one of embodiments 45-48 and/or a second compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

52. A compound of any one of embodiments 45-48 for use in blocking transmission of a Plasmodium parasite in a mammal in need of therefore, optionally for use in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

53. A compound of any one of embodiments 45-48 for use in killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage, optionally for use in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.

54. A method of synthesizing a compound of formula 101, comprising the steps of:

(a) providing a compound of formula 100:

wherein X² is F, Cl, Br, or I,

V is CR¹⁰⁴ or N,

W is CR¹⁰⁵ or N,

Y is CR¹⁰⁶ or N,

Z is CR¹⁰⁷ or N, wherein optionally Y and Z,

wherein R¹⁰⁴-R¹⁰⁷ are independently hydrogen, optionally substituted alkyl, optionally substituted aryl, or optionally substituted heteroaryl, or when Y is CR¹⁰⁶ and Z is CR¹⁰⁷, R¹⁰⁶ and R¹⁰⁶, taken together with the carbons to which they are bound, form an optionally substituted fused 5- to 8-membered carbocyclic, aryl, heterocyclyl, or heteroaryl ring, wherein the heterocyclyl contains one or more atoms selected from N, O, and S,

wherein at least one of V, W, Y, and Z is N,

wherein R¹⁰⁰ is alkyl or aryl, and

wherein R¹⁰¹ is hydrogen, alkyl, or aryl,

(b) reacting the compound of formula 100 with an amine of the formula: R¹⁰²NH₂ to give a compound of the formula 101:

wherein R¹⁰² is optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, optionally substituted arylalkyl, optionally substituted heterocyclylalkyl, optionally substituted cycloalkylalkyl, or optionally substituted heteroarylalkyl.

55. The method of embodiment 54, wherein the reacting step (b) is conducted in the presence of an acid.

56. The method of embodiment 55, wherein the acid is selected from hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, boric acid, hydrofluoric acid, hydrobromic acid, perchloric acid, formic acid, acetic acid, propanoic acid, and trifluoroacetic acid.

57. The method of any one of embodiments 54-56, wherein the reacting is conducted in an aliphatic alcohol solvent.

The following examples further illustrate the invention but, of course, should not be construed as in any way limiting its scope.

Materials and Methods

Cell culture. Asexual parasites of P. falciparum strain 3D7 were cultured as described previously [Trager W, et al., Journal of Parasitology 2005, 91: 484-486]. Stage III-V gametocytes were selected and enriched with 3-day treatment with 50 mM N-acetylglucosamine (NAG) and the following Percoll density gradient centrifugation after gametocyte production [Tanaka T Q, et al., Molecular and Biochemical Parasitology, 2011, 177: 160-163]. Gametocytes of HB3 and Dd2 strains were produced and then set up for assay in a similar process. HepG2 cells (ATCC, cat. no. 77400) were cultured in 175-cm² tissue culture flasks with 30 ml growth medium at 37° C. in a 5% CO₂ humidified atmosphere. Growth medium was made with Dulbecco's Modified Eagle Medium with 10% fetal bovine serum (FBS). Growth medium was replaced every other day and cells were passed at 75% confluence.

Compound library and gametocyte assay screen. The approved drug library was collected with 4,265 compounds from traditional chemical suppliers, specialty collections, pharmacies and custom synthesis [Huang R et al., Science Translational Medicine, 2011, 3: 80ps16] that included 49% drugs approved for human or animal use by the US Food and Drug Administration (FDA), 23% approved in Canada/UK/EU/Japan, and the remaining 28% either in clinical trials or research tool compounds. The Malaria Box contained 400 drugs or tool compounds with the confirmed activities on blood-staged P. falciparum and assessed cytotoxicity against mammalian cells [Gamo F J et al., Nature, 2010 465: 305-U356; Guiguemde W A et al., Nature, 2010, 465: 311-315]. The MIPE library was an internal collection of 550 kinase inhibitors, which contain approved drugs and drug candidates in preclinical and clinical stages [Mathews L A et al., Journal of Biomolecular Screening, 2012, 17: 1231-1242]. Compounds from all libraries were obtained as powder samples and dissolved in DMSO as 10 mM stock solutions, except several hundreds from the approved drug library that were prepared as 4.47 mM stock solutions due to solubility limitations.

Compound screening experiments were performed as previously described [Tanaka T Q et al., Molecular and Biochemical Parasitology, 2013, 3188: 20-25]. Briefly, 2.5 μl/well incomplete medium was dispensed into each well of 1,536-well plates using the Multidrop Combi followed by 23 nl compound transferring using the NX-TR Pintool (WAKO Scientific Solutions, San Diego, Calif.). 2.5 μl/well of gametocytes was dispensed with a seeding density of 20,000 cells/well using the Multidrop Combi. The assay plates were incubated for 72 h at 37° C. with 5% CO₂. After addition of 5 μl/well of 2× AlamarBlue dye (Life Technologies, cat. no. DAL1100), the plates were incubated for 24 h at 37° C. with 5% CO₂ and were read in a fluorescence detection mode (Ex=525 nm, Em=598 nm) on a ViewLux plate reader (PerkinElmer).

Small molecule pull-down. Affinity matrix: To make a bead-connected affinity probe of Torin 2, a tetraethylene glycol linker was attached to 1-(piperazin-1-yl)propan-1-one of HWW030 and then coupled to Affi-Gel 10 resin (Bio-Rad Laboratories, cat. no. 153-6046) under mild basic conditions to afford Torin 2 matrix (T2M). See detailed version in Example 7. Torin 1 was similarly immobilized to resin and used as a negative control (TIM). The resultant affinities probes were incubated with gametocyte lysates, the bound proteins were eluted from resin by boiling in SDS-PAGE sample loading buffer. The eluted fractions were separated by SDS-PAGE and visualized by silver staining. RBC infected with gametocytes (3D7 strain: Stage III-V) were washed 3 times with PBS and then lysed by 0.05% saponin treatment in PBS for 5 min at room temperature. The prepared gametocytes were washed 3 times with PBS and frozen at −80° C. The affinity precipitation experiment was processed as previously described [Zhang Q et al., Proceedings of the National Academy of Sciences of the United States of America, 2007, 104: 7444-7448; Arastu-Kapur S et al., Nature Chemical Biology, 2008, 4: 203-213]. The frozen samples were lysed with homogenization buffer (60 mM glycerophosphate, 15 mM p-nitrophenyl phosphate, 25 mM MOPS (pH 7.2), 15 mM EGTA, 15 mM MgCl2, 1 mM DTT, protease inhibitors (Roche Diagnostics, cat. no. 11836170001), and 0.5% Nonidet P-40). Cell lysates were centrifuged at 16,000×g for 20 min at 4° C., and the supernatant was collected. Protein concentration in the supernatant was determined by using a BCA protein assay kit (Pierce Chemical, cat. no. 23225). The lysate (0.5 mg) was then added to the packed affinity matrix, and bead buffer (50 mM Tris HCl (pH 7.4), 5 mM NaF, 250 mM NaCl, 5 mM EDTA, 5 mM EGTA, protease inhibitors, and 0.1% Nonidet P-40) was added to a final volume of 1 ml. After rotating at 4° C. for 2 h, the mixture was centrifuged at 16,000×g for 2 min at 4° C., and the supernatant was removed. The affinity matrix was then washed (six times) with cold bead buffer and eluted by boiling with SDS-PAGE sample loading buffer at 95° C. for 5 min. Supernatants were separated on a 10% Bis-Tris gel (Life Technologies, cat. no. NP0315BOX) and visualized by silver staining using a Pierce Silver Stain Kit for Mass Spectrometry (Pierce Chemical, cat. no. 24600).

DARTS (drug affinity responsive target stability). The 3D7 gametocytes were lysed with M-PER supplemented with protease and phosphatase inhibitors as previously described [Lomenick B et al., Proceedings of the National Academy of Sciences of the United States of America, 2009, 106: 21984-21989]. After centrifugation at 16,000×g for 20 min, protein concentration in the supernatant was quantified and 2 μg/μl proteins were treated with 600 nM of Torin 2 or 600 nM of Torin 1 for 2 h at room temperature. The samples were treated with 46 μg/ml pronase (Sigma-Aldrich, cat. no. P6911) for 30 min at room temperature. The digestion was stopped by adding the SDS-PAGE sample loading buffer and boiled at 70° C. for 10 min. The samples were separated on a 10% Bis-Tris gel and visualized by silver staining.

Malaria Mouse Model. Plasmodium berghei ANKA (Pb) parasites were maintained by serial passage by intraperitoneal (i.p.) injection in outbred mice. Two days before feeding, female mice were infected i.p. with 200-400 μl whole blood from a Pb-infected mouse with >10% parasitemia. On the day of feeding, the mice were checked for exflagellation and injected intravenously (i.v.) with drug vehicle alone (10% N-methylpyrrolidnone, 40% PEG 400 in water), or (a) 2-4 mg/kg Torin 2 (one or two doses), (b) 8 mg/kg NVP-AUY922 (two doses), or (c) 8 mg/kg Alvespimycin (two doses). Two hours post treatment, mice were anesthetized and Anopheles stephensi mosquitoes were allowed to feed on infected mice for 15 minutes. Parasitemia, gametocytemia, and presence of exflagellation were examined as described previously [Blagborough A M et al., Nature Communications, 2013, 4: 1812]. Mosquitoes were maintained on 5% (w/v) glucose at 19° C. and 80% relative humidity. At day 10 post feeding, mosquito midguts were dissected and transmission was measured by staining mosquito midguts with 0.2% mercurochrome and counting the numbers of oocysts per midgut.

Data analysis. The primary screen data was analyzed using customized software developed internally [Wang Y et al., Current Chemical Genomics 2010, 4: 57-66]. IC₅₀ values were calculated using the Prism software (Graphpad Software, Inc. San Diego, Calif.). Data were presented as means±SEM with n=3 independent experiments.

General materials and methods for chemical synthesis. All commercially available reagents, compounds, and solvents were purchased and used without further purification. 9-Bromo-1-(3-(trifluoromethyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one was prepared according to the method described by Liu and coworkers (Liu, Q. et. al., J. Med. Chem., 2011, 1473-1480). Column chromatography on silica gel was performed on Biotage KPSil pre-packed cartridges using the Biotage SP-1 automated chromatography system. Reverse phase column chromatography was performed on RediSep preparative C-18 column using the Teledyne ISCO combiflash Rf system. ¹H spectra were recorded using an Inova 400 MHz spectrometer (Varian). Samples were analyzed on an Agilent 1200 series LC/MS. Method A used an Enomenex Kinetex 1.7 micron column and a flow rate of 1.1 mL/min. The mobile phase was a mixture of acetonitrile and H₂O each containing 0.05% trifluoroacetic acid. A gradient of 4% to 100% acetonitrile over 4 minutes was used during analytical analysis. Method B used a Zorbax™ Eclipse XDB-C18 reverse phase (5 micron, 4.6×150 mm) column and a flow rate of 1.1 mL/min. The mobile phase was a mixture of acetonitrile and H₂O each containing 0.05% trifluoroacetic acid. A gradient of 5% to 100% acetonitrile over 8 minutes was used during analytical analysis.

Example 1

This example demonstrates an In vitro drug activity on gametocytes.

Stage III-V gametocytes (blood stage P. falciparum parasites) were enriched with treatment with 50 mM N-acetylglucosamine (NAG) and Percoll density gradient centrifugation as described previously¹. Briefly, 2.5 μl/well incomplete medium was dispensed into each well of 1,536-well plates using the Multidrop Combi followed by 23 nl compound transfer using the NX-TR Pintool (WAKO Scientific Solutions, San Diego, Calif.). Then, 2.5 μl/well of gametocytes was dispensed with a seeding density of 20,000 cells/well using the Multidrop Combi. The assay plates were incubated for 72 h at 37° C. with 5% CO₂. After addition of 5 μl/well of 2× AlamarBlue dye (Life Technologies, cat. no. DAL1100), the plates were incubated for 24 h at 37° C. with 5% CO₂ and then were read in a fluorescence detection mode (Ex=525 nm, Em=598 nm) on a ViewLux plate reader (PerkinElmer).

Example 2

This example demonstrates In vitro drug activity on asexual parasites in accordance with an embodiment of the invention.

Asexual parasites of P. falciparum strain 3D7 were cultured as described previously (Trager, W. et al., J. Parasitol. 2005, 91(3): 484-486). Drug activity on asexual stage parasites was tested using a SYBR Green assay as described previously (Eastman, R. T. et al., Antimicrob. Agents Chemother. 2013, 57(1): 425-435; Smilkstein, M. et al., Antimicrob. Agents Chemother. 2004, 48(5): 1803-1806). Briefly, parasites were diluted to 0.5% parasitemia in complete culture medium with 2% hematocrit and drugs diluted in DMSO (<0.5%) and were loaded into a 96-well plate (200 μl/well). No drug and RBC alone wells were included as positive and background controls, respectively, and each testing condition was examined in duplicated. After 72 h incubation under the standard culture condition and a freeze-thaw lysis step at −80° C. and room temperature, 100 μl/well of lysis buffer containing SYBR Green I was added to the parasite culture and incubated for 30 min at room temperature. The fluorescence of each well was measured at 520 nm following excitation at 490 nm using a FLUOstar Optima™ microplate reader (BMG Labtech).

Example 3

This example demonstrates a synthesis of compounds, in accordance with an embodiment of the invention.

A general procedure for the synthesis of compound 2 is shown in Scheme 1.

Aldehydes 1 were prepared using a reported procedure (J. Med. Chem. 2011, 54(5): 1473-1480). A solution of 1 (300 μmole) in 3 mL of THF were added 300 μL of Et₂NiPr and R′CH₂COCl (3000 μmole). The mixture was heated in a microwave between 100 to 150° C. for 15 min. The crude product was purified by column chromatography on silica gel using dichloromethane in methanol (0-20%) as eluent to give 1′. A mixture of 1′ (1.0 equiv), boronic acid or boronic acid pinacol ester (3.0 equiv), tetrakis(triphenylphosphine)palladium (0.05 equiv), DMF (1.5 mL) and saturated NaHCO₃ aqueous solution (0.5 mL) was charged in a microwave vial. Nitrogen was bubbled through the mixture for 3 min. The vial was capped and heated in a microwave at 120-150° C. for 15 min. The reaction mixture was filtered through a plug of Celite and the filtrate was purified by reverse phase column chromatography using acetonitrile (containing 0.1% TFA)/water (containing 0.1% TFA) as an eluent to give 2.

9-(4-chlorophenyl)-1-(4-(morpholine-4-carbonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 496.1 (MH)⁺, t_(R)=4.712, UV₂₅₄=100%

9-(4-((dimethylamino)methyl)phenyl)-1-(4-(morpholine-4-carbonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 519.2 (MH)⁺, t_(R)=3.462, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 466.1 (MH)⁺, t_(R)=4.659, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-cyclopropyl-3-methylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 480.1 (MH)⁺, t_(R)=4.765, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-(2,2,2-trifluoroethyl)benzamide

White solid, isolated yield: 18.3%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.42 (t, J=6.2 Hz, 1H), 9.17 (s, 1H), 8.35 (d, J=9.5 Hz, 1H), 8.23-8.17 (m, 2H), 8.12 (dd, J=8.6, 0.5 Hz, 1H), 8.01 (dd, J=8.7, 2.0 Hz, 1H), 7.73-7.67 (m, 2H), 7.35-7.28 (m, 2H), 7.09-7.05 (m, 3H), 6.95 (d, J=9.4 Hz, 1H), 4.30-4.16 (m, 2H); LC/MS (Method B): (electrospray +ve), m/z 508.0 (MH)⁺, t_(R)=4.992, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-(2-(dimethylamino)ethyl)benzamide

Off-white solid, isolated yield: 28.4%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.19 (s, 1H), 8.95 (t, J=5.7 Hz, 1H), 8.36 (d, J=9.5 Hz, 1H), 8.19-8.10 (m, 3H), 8.02 (dd, J=8.7, 1.9 Hz, 1H), 7.73-7.67 (m, 2H), 7.38-7.31 (m, 2H), 7.14-7.08 (m, 2H), 7.06 (dd, J=1.9, 0.5 Hz, 1H), 6.96 (d, J=9.4 Hz, 1H), 3.73 (q, J=6.0 Hz, 2H), 3.34 (q, J=6.0 Hz, 2H), 2.90 (d, J=4.8 Hz, 7H); LC/MS (Method B): (electrospray +ve), m/z 497.3 (MH)⁺, t_(R)=3.667, UV₂₅₄=100%

N-cyclopropyl-4-(9-(4-fluorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)benzamide

Off-white solid, isolated yield: 19.25%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.16 (s, 1H), 8.70 (d, J=4.4 Hz, 1H), 8.34 (d, J=9.5 Hz, 1H), 8.11 (dd, J=8.5, 6.3 Hz, 3H), 7.99 (dd, J=8.7, 2.0 Hz, 1H), 7.65-7.60 (m, 2H), 7.16-7.08 (m, 4H), 6.97 (d, J=2.3 Hz, 1H), 6.95 (d, J=9.4 Hz, 1H), 3.02-2.94 (m, 1H), 0.79 (td, J=7.0, 4.6 Hz, 2H), 0.69-0.59 (m, 2H); LC/MS (Method B): (electrospray +ve), m/z 450.1 (MH)⁺, t_(R)=4.372, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-(1-(2-(dimethylamino)ethyl)piperidin-4-yl)benzamide

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-(1-(2-hydroxyethyl)azetidin-3-yl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 525.2 (MH)+, t_(R)=2.666, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(4-(4-(2-(dimethylamino)ethyl)piperazine-1-carbonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 566.3 (MH)+, t_(R)=2.731, UV₂₅₄=100%

4-(9-(4-chlorophenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-(pyrrolidin-3-yl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 495.2 (MH)⁺, t_(R)=2.713, UV₂₅₄=100%

4-(9-(4-(1-aminocyclopropyl)phenyl)-2-oxobenzo[h][1,6]naphthyridin-1(2H)-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 487.3 (MH)⁺, t_(R)=2.264, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(1-oxoisoindolin-5-yl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 438.1 (MH)⁺, t_(R)=4.145, UV₂₅₄=100%

9-(4-((dimethylamino)methyl)phenyl)-1-(1-oxoisoindolin-5-yl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 461.2 (MH)⁺, t_(R)=3.149, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(2-methyl-1-oxoisoindolin-5-yl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 452.1 (MH)⁺, t_(R)=4.309, UV₂₅₄=100%

9-(4-((dimethylamino)methyl)phenyl)-1-(1-oxo-1,2,3,4-tetrahydroisoquinolin-6-yl)benzo[h][1,6]naphthyridin-2(1H)-one

Isolated yield: 30.1%. LC/MS (Method B): (electrospray +ve), m/z 475.1 (MH)⁺, t_(R)=3.257, UV₂₅₄=100%

1-(2-(3-aminopropyl)-1-oxoisoindolin-5-yl)-9-(4-chlorophenyl)benzo[h][1,6]naphthyridin-2(1H)-one

Off-white solid, isolated yield: 24.83%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.19 (s, 1H), 8.36 (d, J=9.5 Hz, 1H), 8.12 (dd, J=8.7, 0.5 Hz, 1H), 8.00 (dd, J=8.7, 2.0 Hz, 1H), 7.96 (dd, J=8.0, 0.6 Hz, 1H), 7.89 (dd, J=1.8, 0.7 Hz, 1H), 7.74 (s, 4H), 7.64 (dd, J=8.0, 1.8 Hz, 1H), 7.30-7.24 (m, 2H), 7.05-7.00 (m, 2H), 6.97 (d, J=9.5 Hz, 1H), 6.91 (dd, J=2.0, 0.6 Hz, 1H), 4.75-4.55 (m, 2H), 3.83 (dt, J=14.3, 7.3 Hz, 1H), 3.63 (dt, J=13.6, 6.5 Hz, 1H), 2.89 (s, 2H), 2.02-1.91 (m, 2H); LC/MS (Method B): (electrospray +ve), m/z 495.0 (MH)⁺, t_(R)=3.783, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 461.1 (MH)⁺, t_(R)=4.389, UV₂₅₄=100%

9-(4-chloro-3-hydroxyphenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 477.1 (MH)+, t_(R)=4.178, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(4-(ethylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

9-(3-amino-4-chlorophenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 476.1 (MH)⁺, t_(R)=4.331, UV₂₅₄=100%

9-(4-fluorophenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 445.1 (MH)⁺, t_(R)=4.478, UV₂₅₄=100%

9-(4-((dimethylamino)methyl)phenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 484.2 (MH)+, t_(R)=3.264, UV₂₅₄=100%

9-(4-chloro-3-methoxyphenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

Off-white solid, isolated yield: 38.6%. LC/MS (Method B): (electrospray +ve), m/z 491.1 (MH)+, t_(R)=4.866, UV₂₅₄=100%

2-chloro-5-(1-(4-(methylsulfonyl)phenyl)-2-oxo-1,2-dihydrobenzo[h][1,6]naphthyridin-9-yl)benzonitrile

Yellow solid, isolated yield: 9.67%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.22 (s, 1H), 8.37 (d, J=9.5 Hz, 1H), 8.23 (dt, 2H), 8.18-8.14 (m, 2H), 8.10 (dd, J=8.7, 1.9 Hz, 1H), 7.86 (dt, 2H), 7.67 (d, J=8.5 Hz, 1H), 7.00 (d, J=1.9 Hz, 1H), 6.98 (d, J=9.4 Hz, 1H), 6.92 (dd, J=8.6, 2.3 Hz, 1H), 3.36 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 486.1 (MH)⁺, t_(R)=4.634, UV₂₅₄=100%

1-(4-(methylsulfonyl)phenyl)-9-(4-(pyrrolidin-1-ylmethyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

Off-white solid, isolated yield: 2.405%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.17 (s, 1H), 8.35 (d, J=9.5 Hz, 1H), 8.28-8.22 (m, 2H), 8.12 (d, J=8.6 Hz, 1H), 8.01 (dd, J=8.7, 1.9 Hz, 1H), 7.88-7.83 (m, 2H), 7.34 (d, J=7.7 Hz, 2H), 7.12-7.09 (m, 1H), 7.02 (d, J=7.8 Hz, 2H), 6.96 (d, J=9.4 Hz, 1H), 2.44 (s, 5H), 1.72 (s, 5H); LC/MS (Method B): (electrospray +ve), m/z 510.4 (MH)⁺, t_(R)=3.302, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(2-methyl-4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

Yellow solid, isolated yield: 2.89%. ¹H NMR (400 MHz, DMSO-d₆) 9.21 (s, 1H), 8.40 (d, J=9.5 Hz, 1H), 8.23 (dd, J=1.9, 0.7 Hz, 1H), 8.15 (dd, J=8.6, 0.5 Hz, 1H), 8.11-8.03 (m, 2H), 7.77-7.73 (m, 1H), 7.49-7.44 (m, 2H), 7.13-7.06 (m, 2H), 7.05-7.03 (m, 1H), 7.00 (d, J=9.5 Hz, 1H), 3.36 (s, 3H), 2.12 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 475.1 (MH)⁺, t_(R)=4.966, UV₂₅₄=100%

9-(4-chlorophenyl)-1-(4-(methylsulfonyl)-3-(piperazin-1-ylmethyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

LC/MS (Method B): (electrospray +ve), m/z 559.1 (MH)⁺, t_(R)=4.318, UV₂₅₄=100%

9-(4-(1-aminocyclopropyl)phenyl)-1-(4-(methylsulfonyl)phenyl)benzo[h][1,6]naphthyridin-2(1H)-one

Off-white solid, isolated yield: 3.24%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.19 (s, 1H), 8.36 (d, J=9.5 Hz, 1H), 8.26-8.20 (m, 2H), 8.14 (dd, J=8.6, 0.5 Hz, 1H), 8.03 (dd, J=8.7, 1.9 Hz, 1H), 7.89-7.84 (m, 2H), 7.46-7.41 (m, 2H), 7.15-7.09 (m, 2H), 7.05 (dd, J=2.0, 0.6 Hz, 1H), 6.97 (d, J=9.4 Hz, 1H), 3.34 (s, 4H), 1.38-1.32 (m, 2H), 1.22-1.16 (m, 2H); LC/MS (Method B): (electrospray +ve), m/z 482.1 (MH)⁺, t_(R)=3.459, UV₂₅₄=100%

Example 4

This example demonstrates a synthesis of compounds, in accordance with an embodiment of the invention.

A reaction scheme for the synthesis of compounds 13 and 14 is shown in Scheme 2.

8-(4-chloro-3-hydroxyphenyl)-3-methyl-1-(4-(morpholine-4-carbonyl)phenyl)-1H-imidazo[4,5-c]quinolin-2(3H)-one

Off-white solid, isolated yield: 24%. LC/MS (Method B): (electrospray +ve), m/z 515.2 (MH)⁺, t_(R)=3.995, UV₂₅₄=100%

4-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-N,N-dimethylbenzamide

Off-white solid, isolated yield: 18%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.16 (s, 1H), 8.17 (d, J=8.9 Hz, 1H), 8.01 (d, J=8.9 Hz, 1H), 7.78-7.72 (m, 4H), 7.46 (d, J=8.5 Hz, 2H), 7.38 (d, J=8.6 Hz, 2H), 7.23 (s, 1H), 3.64 (s, 3H), 3.08 (s, 3H), 2.98 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 457.1 (MH)⁺, t_(R)=4.153, UV₂₅₄=100%

4-(8-(4-chloro-3-hydroxyphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-N-cyclopropyl-N-methylbenzamide

Off-white solid, isolated yield: 18%. ¹H NMR (400 MHz, DMSO-d₆) δ 10.28 (s, 1H), 9.12 (s, 1H), 8.15 (d, J=8.9 Hz, 1H), 7.82 (d, J=8.6 Hz, 1H), 7.79 (d, J=8.3 Hz, 2H), 7.73 (d, J=8.3 Hz, 2H), 7.36 (s, 1H), 7.31 (d, J=8.2 Hz, 1H), 7.29 (s, 1H), 6.95 (s, 1H), 6.76 (d, J=8.1 Hz, 1H), 3.63 (s, 3H), 3.02 (s, 3H), 2.93-2.87 (m, 1H), 0.33 (d, J=70.3 Hz, 3H); LC/MS (Method B): (electrospray +ve), m/z 499.2 (MH)⁺, t_(R)=4.162, UV₂₅₄=100%

4-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-N-(3-(dimethylamino)propyl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 514.1 (MH)⁺, t_(R)=3.492, UV₂₅₄=100%

N-cyclopropyl-4-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 453.2 (MH)⁺, t_(R)=3.894, UV₂₅₄=100%

3-(8-(3-(dimethylamino)prop-1-yn-1-yl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 382.1 (MH)⁺, t_(R)=2.916, UV₂₅₄=100%

3-(8-(4-chlorophenyl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 397.1 (MH)⁺, t_(R)=4.452, UV₂₅₄=100%

3-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 411.1 (MH)⁺, t_(R)=4.626, UV₂₅₄=100%

3-(8-(4-chlorophenyl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 22%. ¹H NMR (400 MHz, DMSO-d₆) δ 12.16 (s, 1H), 8.94 (s, 1H), 8.28 (d, J=1.8 Hz, 1H), 8.16 (d, J=8.9 Hz, 1H), 8.12 (dd, J=8.0, 1.8 Hz, 1H), 7.99 (dd, J=9.0, 2.1 Hz, 1H), 7.84 (d, J=8.1 Hz, 1H), 7.52-7.47 (m, 2H), 7.37-7.33 (m, 2H), 6.98 (d, J=2.0 Hz, 1H), 2.21 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 411.1 (MH)⁺, t_(R)=4.666, UV₂₅₄=100%

3-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 9%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.16 (s, 1H), 8.26 (d, J=1.8 Hz, 1H), 8.17 (d, J=8.9 Hz, 1H), 8.13 (dd, J=8.0, 1.7 Hz, 1H), 7.98 (dd, J=8.9, 2.0 Hz, 1H), 7.85 (d, J=8.0 Hz, 1H), 7.52-7.48 (m, 2H), 7.37-7.33 (m, 2H), 6.98 (d, J=2.0 Hz, 1H), 3.65 (s, 3H), 2.20 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 425.1 (MH+), t_(R)=4.847, UV₂₅₄=100%

3,3′-(3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinoline-1,8-diyl)dibenzonitrile

Off-white solid, isolated yield: 37%. LC/MS (Method B): (electrospray +ve), m/z 402.1 (MH)⁺, t_(R)=4.011, UV₂₅₄=100%

3-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

Off-white solid, isolated yield: 20%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.07 (s, 1H), 8.33 (s, 1H), 8.18-8.12 (m, 2H), 8.09 (d, J=8.2 Hz, 1H), 7.93 (t, J=8.0 Hz, 2H), 7.42 (dd, J=8.7, 5.4 Hz, 2H), 7.27 (t, J=8.7 Hz, 2H), 7.16 (s, 1H), 3.62 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 395.1 (MH)⁺, t_(R)=4.262, UV₂₅₄=100%

3-(8-(4-chloro-3-methoxyphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

Off-white solid, isolated yield: 7%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.03 (s, 1H), 8.30 (d, J=2.0 Hz, 1H), 8.16 (d, J=7.8 Hz, 1H), 8.12 (d, J=8.9 Hz, 1H), 8.07 (dd, J=7.6, 1.9 Hz, 1H), 7.93 (dd, 1H), 7.90 (t, J=8.0 Hz, 1H), 7.45 (d, J=8.2 Hz, 1H), 7.17 (d, J=2.0 Hz, 1H), 7.01 (dd, J=8.2, 2.0 Hz, 1H), 6.96 (d, J=2.0 Hz, 1H), 3.85 (s, 3H), 3.60 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 441.1 (MH)⁺, t_(R)=4.583, UV₂₅₄=100%

3-(8-(4-chlorophenyl)-3-(2-hydroxyethyl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

Off-white solid, isolated yield: 12%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.15 (s, 1H), 8.33 (d, J=2.0 Hz, 1H), 8.17 (t, J=8.1 Hz, 2H), 8.10 (dd, J=7.7, 1.8 Hz, 1H), 7.99-7.91 (m, 2H), 7.52-7.48 (m, 2H), 7.42-7.39 (m, 2H), 7.21 (d, J=2.0 Hz, 1H), 4.19 (t, J=5.3 Hz, 2H), 3.81 (d, J=5.4 Hz, 2H), 3.48 (s, 1H); LC/MS (Method B): (electrospray +ve), m/z 441.1 (MH)⁺, t_(R)=4.389, UV₂₅₄=100%

3-(3-acetyl-8-(4-chlorophenyl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

Off-white solid, isolated yield: 8%. LC/MS (Method B): (electrospray +ve), m/z 439.1 (MH)⁺, t_(R)=5.215, UV₂₅₄=100%

5-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 495.2 (MH)⁺, t_(R)=3.337, UV₂₅₄=100

3-(8-(4-chloro-3-methoxyphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 17%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.12 (s, 1H), 8.26 (d, J=1.8 Hz, 1H), 8.16 (d, J=8.9 Hz, 1H), 8.12 (dd, J=8.0, 1.8 Hz, 1H), 7.99 (dd, J=8.9, 2.1 Hz, 1H), 7.83 (d, J=8.0 Hz, 1H), 7.48 (d, J=8.2 Hz, 1H), 7.02 (d, J=2.0 Hz, 1H), 7.02-6.99 (m, 1H), 6.93 (d, J=2.1 Hz, 1H), 3.88 (s, 3H), 3.65 (s, 3H), 2.19 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 455.1 (MH)⁺, t_(R)=4.581, UV₂₅₄=100%

5-(1-(5-cyano-2-methylphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-8-yl)-2-fluorobenzonitrile

Off-white solid, isolated yield: 19%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.10 (s, 1H), 8.24 (d, J=1.7 Hz, 1H), 8.16 (d, J=8.9 Hz, 1H), 8.08 (dd, J=8.0, 1.8 Hz, 1H), 7.97 (dd, J=8.9, 2.1 Hz, 1H), 7.88 (dd, J=6.1, 2.3 Hz, 1H), 7.84 (d, J=8.1 Hz, 1H), 7.61 (d, J=9.0 Hz, 1H), 7.21-6.95 (m, 1H), 6.94 (s, 1H), 3.65 (s, 3H), 2.19 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 434.2 (MH)⁺, t_(R)=4.248, UV₂₅₄=100%

3-(8-(4-fluoro-3-methoxyphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 19%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.13 (s, 1H), 8.26 (d, J=1.8 Hz, 1H), 8.16 (d, J=8.9 Hz, 1H), 8.12 (dd, J=8.0, 1.8 Hz, 1H), 7.99 (dd, J=9.0, 2.0 Hz, 1H), 7.84 (d, J=8.1 Hz, 1H), 7.27 (dd, J=11.3, 8.4 Hz, 1H), 7.00-6.93 (m, 3H), 3.86 (s, 3H), 3.65 (s, 3H), 2.20 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 439.1 (MH)⁺, t_(R)=4.290, UV₂₅₄=100%

2-chloro-5-(1-(5-cyano-2-methylphenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-8-yl)benzonitrile

Off-white solid, Isolated yield: 15%. LC/MS (Method B): (electrospray +ve), m/z 450.1 (MH)⁺, t_(R)=4.462, UV₂₅₄=100%

3-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 23%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.15 (s, 1H), 8.26 (d, J=1.8 Hz, 1H), 8.16 (d, J=8.9 Hz, 1H), 8.12 (dd, J=8.0, 1.8 Hz, 1H), 7.98 (d, J=2.1 Hz, 1H), 7.85 (d, J=8.0 Hz, 1H), 7.39-7.35 (m, 2H), 7.27 (t, J=8.8 Hz, 2H), 6.97 (d, J=2.1 Hz, 1H), 3.65 (s, 3H), 2.21 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 409.1 (MH)⁺, t_(R)=4.279, UV₂₅₄=100%

3-(8-(3-cyanophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

Off-white solid, isolated yield: 16%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.09 (s, 1H), 8.26 (d, J=1.8 Hz, 1H), 8.16 (d, J=8.8 Hz, 1H), 8.09 (dd, J=8.0, 1.8 Hz, 1H), 7.99 (dd, J=8.9, 2.1 Hz, 1H), 7.85 (d, J=8.2 Hz, 2H), 7.77 (d, J=1.4 Hz, 1H), 7.63 (dd, 2H), 6.99 (d, J=2.1 Hz, 1H), 3.65 (s, 3H), 2.20 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 416.1 (MH)⁺, t_(R)=4.052, UV₂₅₄=100%

3-(8-(4-(2-aminoethyl)phenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

Off-white solid, isolated yield: 28%. LC/MS (Method B): (electrospray +ve), m/z 420.1 (MH)⁺, t_(R)=3.131, UV₂₅₄=100%

5-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 479.2 (MH)⁺, t_(R)=3.135, UV₂₅₄=100%

5-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methyl-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 509.2 (MH)⁺, t_(R)=3.378, UV₂₅₄=100%

5-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methyl-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 493.3 (MH)⁺, t_(R)=3.195, UV₂₅₄=100%

5-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-(4-(2-hydroxyethyl)piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 539.3 (MH)⁺, t_(R)=3.401, UV₂₅₄=100%

5-(8-(3-cyanophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 486.1 (MH)⁺, t_(R)=3.282, UV₂₅₄=100%

4-methyl-3-(3-methyl-2-oxo-8-(1H-pyrrol-2-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 380.1 (MH)⁺, t_(R)=3.645, UV₂₅₄=100%

4-methyl-3-(3-methyl-8-(1-methyl-1H-pyrrol-2-yl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 394.1 (MH)⁺, t_(R)=4.004, UV₂₅₄=100%

4-methyl-5-(3-methyl-2-oxo-8-(thiophen-2-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 481.1 (MH)⁺, t_(R)=2.962, UV₂₅₄=100%

2-(4-acetylpiperazin-1-yl)-5-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 535.1 (MH)⁺, t_(R)=4.093, UV₂₅₄=100%

2-(4-acetylpiperazin-1-yl)-5-(8-(3-cyanophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 542.1 (MH)⁺, t_(R)=3.954, UV₂₅₄=100%

4-methyl-3-(3-methyl-2-oxo-8-(thiophen-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 397.1 (MH)⁺, t_(R)=3.920, UV₂₅₄=100%

3-(8-(furan-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 381.1 (MH)+, t_(R)=3.845, UV₂₅₄=100%

2-(3-aminoazetidin-1-yl)-5-(8-(3-cyanophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 486.1 (MH)⁺, t_(R)=3.290, UV₂₅₄=100%

5-(8-(3-cyano-4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methyl-2-(piperazin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 518.1 (MH)⁺, t_(R)=3.455, UV₂₅₄=100%

2-(3-aminoazetidin-1-yl)-5-(8-(4-fluorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 479.1 (MH)⁺, t_(R)=3.380, UV₂₅₄=100%

2-(4-acetylpiperazin-1-yl)-5-(8-(4-chlorophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 551.1 (MH)⁺, t_(R)=4.309, UV₂₅₄=100%

5-(8-(3-cyanophenyl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)-2-((2-(dimethylamino)ethyl)amino)-4-methylbenzonitrile

LC/MS (Method B): (electrospray +ve), m/z 502.1 (MH)⁺, t_(R)=3.348, UV₂₅₄=100%

8-(4-chlorophenyl)-3-methyl-1-(4-(methylsulfonyl)phenyl)-1H-imidazo[4,5-c]quinolin-2(3H)-one

Off-white solid, isolated yield: 12%. ¹H NMR (400 MHz, DMSO-d₆) δ 9.05 (s, 1H), 8.28 (d, J=8.4 Hz, 2H), 8.13 (d, J=8.9 Hz, 1H), 8.00 (d, J=8.4 Hz, 2H), 7.92 (dd, J=8.9, 2.1 Hz, 1H), 7.43-7.36 (m, 4H), 7.11 (d, J=2.0 Hz, 1H), 3.62 (s, 3H), 3.41 (s, 3H); LC/MS (Method B): (electrospray +ve), m/z 464.1 (MH)⁺, t_(R)=4.238, UV₂₅₄=100%

8-(4-chlorophenyl)-1-(4-(methylsulfonyl)phenyl)-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one

LC/MS (Method B): (electrospray +ve), m/z 450.1 (MH)⁺, t_(R)=4.119, UV₂₅₄=100%

Example 5

This example demonstrates a synthesis of compounds, in accordance with an embodiment of the invention.

A representative synthesis is shown in Scheme 3. To a 10 mL microwave tube charged with bromide 1 (2.25 mmol) in acetic acid (5 mL) were added hydrazine 2 (4.5 mmol, 2.0 equiv). The reaction was heated under MW at 180° C. for 25 min, and was monitored by LC-MS. Then the reaction mixture was concentrated in vacuo, and the product was triturated with water to afford the product 3 in 75% yield.

To a 10 mL microwave tube charged with bromide 3 (0.3 mmol in DMF (1.5 mL) were added i-PrOH (0.75 mL) and 0.75 mL aqueous K3PO4 solution (0.8 M, 2 equiv). The mixture was stirred for 5 min. Nitrogen gas was bubbled through the reaction solution for 1-2 min, then Pd(PP3)4 (0.05 equiv) and boronic acid/ester 4 (2 equiv) were added. The mixture was heated in a MW at 150° C. for 10 min. The mixture was filtered through Celite and the filtrate was concentrated under vacuum. The crude was treated with 50% TFA in CH₂Cl₂ at room temperature for 2 h, then the solvent was removed under vacuum, and the crude was purified by silica gel chromatograph with DCM/10% NH₄OH in MeOH.

(4-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)phenyl)(pyrrolidin-1-yl)methanone

LC/MS (Method B): (electrospray +ve), m/z 467.2 (MH)⁺, t_(R)=4.705, UV₂₅₄=100%

4-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 453.1 (MH)⁺, t_(R)=4.536, UV₂₅₄=100%

4-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)-N-(3-(dimethylamino)propyl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 498.2 (MH)⁺, t_(R)=3.795, UV₂₅₄=100%.

(4-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)phenyl)(morpholino)methanone

LC/MS (Method B): (electrospray +ve), m/z 483.1 (MH)⁺, t_(R)=4.350, UV₂₅₄=100%.

(4-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)-2-(trifluoromethyl)phenyl) (morpholino)methanone

LC/MS (Method B): (electrospray +ve), m/z 551.1 (MH)⁺, t_(R)=4.830, UV₂₅₄=100%

N-cyclopropyl-3-(8-(4-((dimethylamino)methyl)phenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)benzamide

LC/MS (Method B): (electrospray +ve), m/z 476.2 (MH)⁺, t_(R)=3.265, UV₂₅₄=100%.

(3-(8-(4-((dimethylamino)methyl)phenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)phenyl)(morpholino)methanone

LC/MS (Method B): (electrospray +ve), m/z 506.2 (MH)⁺, t_(R)=3.193, UV₂₅₄=100%.

3-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 453.2 (MH)⁺, t_(R)=4.659, UV₂₅₄=100%.

(3-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)phenyl)(morpholino)methanone

LC/MS (Method B): (electrospray +ve), m/z 483.2 (MH)⁺, t_(R)=4.503, UV₂₅₄=100%.

4-(8-(4-chlorophenyl)-1H-pyrrolo[3,2-c]quinolin-1-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 438.2 (MH)⁺, t_(R)=5.765, UV₂₅₄=100%.

3-(8-(6-aminopyridin-3-yl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)-N-cyclopropylbenzamide

LC/MS (Method B): (electrospray +ve), m/z 435.2 (MH)⁺, t_(R)=3.107, UV₂₅₄=100%.

(3-(8-(6-aminopyridin-3-yl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)phenyl)(morpholino)methanone

LC/MS (Method B): (electrospray +ve), m/z 465.2 (MH)⁺, t_(R)=3.061, UV₂₅₄=100%.

3-(8-(4-chlorophenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 395.1 (MH)⁺, t_(R)=5.027, UV₂₅₄=100%.

3-(8-(3-(1-aminocyclopropyl)phenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)benzonitrile

3-(8-(4-((dimethylamino)methyl)phenyl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 418.2 (MH)⁺, t_(R)=3.387, UV₂₅₄=100%.

3-(8-(6-aminopyridin-3-yl)-3-methyl-1H-pyrazolo[4,3-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 377.1 (MH)⁺, t_(R)=3.219, UV₂₅₄=100%.

4-methyl-3-(3-methyl-8-(1-methyl-1H-pyrazol-4-yl)-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 395.2 (MH)⁺, t_(R)=3.376, UV₂₅₄=100%.

4-methyl-3-(3-methyl-2-oxo-8-(1H-pyrazol-4-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 381.1 (MH)⁺, t_(R)=3.100, UV₂₅₄=100%.

1-(4-methoxyphenyl)-3-methyl-8-(thiophen-3-yl)-1,3-dihydro-2H-imidazo[4,5-c]quinolin-2-one

LC/MS (Method B): (electrospray +ve), m/z 388.1 (MH)⁺, t_(R)=4.054, UV₂₅₄=100%.

4-(dimethylamino)-3-(3-methyl-2-oxo-8-(thiophen-2-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 426.1 (MH)⁺, t_(R)=4.021, UV₂₅₄=100%.

4-(dimethylamino)-3-(3-methyl-2-oxo-8-(thiophen-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 426.1 (MH)⁺, t_(R)=4.082, UV₂₅₄=100%.

3-(3-methyl-2-oxo-8-(thiophen-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 383.1 (MH)⁺, t_(R)=3.893, UV₂₅₄=100%.

4-fluoro-3-(3-methyl-2-oxo-8-(thiophen-2-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 401.1 (MH)⁺, t_(R)=4.139, UV₂₅₄=100%.

3-(3-methyl-2-oxo-8-(thiophen-2-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 383.1 (MH)+, t_(R)=4.019, UV₂₅₄=100%.

4-(dimethylamino)-3-(8-(furan-3-yl)-3-methyl-2-oxo-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 410.1 (MH)⁺, t_(R)=3.867, UV₂₅₄=100%.

4-methyl-3-(3-methyl-2-oxo-8-(1H-pyrrol-3-yl)-2,3-dihydro-1H-imidazo[4,5-c]quinolin-1-yl)benzonitrile

LC/MS (Method B): (electrospray +ve), m/z 380.2 (MH)⁺, t_(R)=3.486, UV₂₅₄=100%.

Example 6

This example demonstrates the gametocytocidal activity and activity against asexual parasites in accordance with an embodiment of the invention.

Compounds were screened against gametocytes and asexual parasites as described in Examples 1 and 2. The results are set forth in Tables 1-8.

TABLE 1 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 1 88.85 27.95 643.6 — — — 2 427.5 291 4316 >30.0 >64.0 1040.6 3 264.3 111.4 12820 12.1 <0.1 621.2 4 54.99 14.92 12820 9.6 <1 21 5 507.5 N/A 3557 — 7.03 725.27 6 101.7 N/A 100000 — 0.02 ND 7 25.07 13.05 1433 — >66.00 — 8 139.3 60.32 1305 14.5 <1 <1 9 2570 667.7 50000 10.9 58.6 1492.3 10 469.2 72.19 2077 18 >66 0 11 54.99 14.92 12820 9.6 <1 21 12 125.5 55.82 50000 — — — 13 135.1 80.36 50000 0 0 0 14 425 113 114300 — — — 15 521.2 730.4 50000 — — — 16 69.39 23.74 6175 — — — 17 — — — — — — 18 — — — — — — 19 — — — — — — 20 22.59 11.65 42.71 >30.0 <1.0 505.6 21 17.02 13.79 232.5 12.7 1.1 606 22 114.7 34.56 101.7 >30.0 <1.0 ND 23 40.35 22.47 182.9 3.5 1.48 58.15 24 23.35 5.876 75.31 19 5.7 547.9

TABLE 2 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 25 117 14.72 6251 >30.0 <1.0 <1.5 26 177.6 37.83 27290 >30.0 1.2 ND 27 107.8 0 178.9 >30.0 <1.0 <1.0 28 95.31 16.17 57.76 >30.0 <1.0 33.9 29 88.48 19.75 119.2 >30.0 <1.0 — 30 336.3 117.9 2902 >30.0 30.4 677.9 31 222 33.47 458 >30.0 <1.0 40.7 32 92.76 85.53 50000 >30.0 <1.0 39.6 33 — 288.4 6606 >30 61 439.2 34 — 59.4 4268 17.1 <1 <1 35 — 75.77 0 >30 4.3 35 36 — 187.7 1083 >30 5.9 6.5 37 — — — — — — 38 187.1 219.7 1116 12.7 <1.0 32.5 39 426 413.7 469.2 1.7 <1.0 — 40 51.73 14.6 174.3 >30.0 <1.0 <1.0 41 89.41 23.12 189.5 6.3 <1.0 ND 42 118.5 37.36 254.7 4.9 <1.0 97.4 43 Inactive 43850 1642 >30.0 >74.0 <1.0 44 135.4 31.17 86.36 4.6 <1.0 34.8 45 208.8 77.83 32.8 8.9 <1.0 144.4 46 259.7 62.51 1275 8.3 <1.0 ND 47 377.3 66.37 399.4 >30.0 2.1 <2.9 48 560.9 200.2 1315 >30.0 <1.0 <1.0 49 1994 3192 448.2 4.3 <1.0 235.8 50 1826 932.4 2698 3 2.7 164.4 51 291.3 202.9 1658 >30.0 4.3 43.1 52 35210 14730 25790 <1.0 57.2 200.8 53 6850 3791 11060 2.8 9.6 115.9 54 393.1 145.9 1678 51.7 >69.00 — 55 5335 N/A 3537 9.45 >71.00 788.14 56 1548 N/A 440.7 >30.00 0.01 716.98 57 839.9 380.6 3371 >30.0 7.9 527.8 58 890.4 N/A 10570 13.5 <0.10 1527.98 59 25.36 9.069 345.5 12.8 <0.10 <1.00 60 1951 N/A 18480 >30.00 21.28 1646.44 61 8.486 211.6 >30 2.3 <1 62 1325 558.5 11610 >30.00 >84.00 — 63 342.1 147.3 4738 — — — 64 1046 488.7 350 >30 1.4 <12.3 65 461 43.08 756.2 2.9 <1 5.4 66 350.4 — 1308 1.4 <1 135 67 1338 — 6117 1.7 <1 1449.1 68 — 131.4 8033 >30 15.7 148.7 69 — 1940 100000 >30 >73 828.3 70 — 20.1 373.9 31.3 1.2 148.4 71 — 56.65 2106 >30 2 133.5 72 — — — — — — 73 — — — — — — 74 — — — — — — 75 — — — — — — 76 — — — — — — 77 294.6 81.67 17.03 >30.0 33.6 <1.0 78 11.57 6.116 10.15 >30.0 <1.0 ND 79 22.81 4.756 32.37 >30.0 <1.0 <11.8 80 213.6 N/A 33.29 7.1 49.68 50.85 81 33.86 N/A 81.93 — 10 5.1 82 166.7 150.4 231.4 — — — 83 74.74 N/A 173.6 >30.00 5.62 58.31 84 — — — — — —

TABLE 3 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 85 77.58 6.895 — 9 <1.0 793.8 86 211.5 177.2 3776 >30.0 61.6 459 87 139.2 24.36 241.1 >30.0 2.6 33.1 88 141.3 56.7 1841 — 0.51 117.49 89 — 52.14 2689 >30 <1 <1 90 — 74.11 19840 >30 13.6 802 91 — 33.02 4415 >30 <1 54.2 92 — 27.2 613.8 >30 13.3 3.9 93 — 27.9 450.7 >30 <1 76.3 94 — 24.92 4188 >30 13.6 1.9 95 — 15.67 1731 >30 54.7 674 96 — 37.29 1291 >30 11.4 7.8 97 — 100.6 1400 >30 7.1 35.8 98 — 19.34 994 >30 0 0 99 130 46.68 — 16.9 <1.0 1054.5 100 45.51 14.78 1531 3.8 <1.0 314.6 101 305.4 107 1780 4.5 <1.0 — 102 116.5 185.8 5191 5 7.5 1083.4 103 71.34 74.26 2997 10.5 6.3 76.1 104 193.7 214.7 2680 1.4 <1.0 388.8 105 94.33 47.9 779.9 5.9 14.8 284.6 106 125.6 63.99 1735 2.1 <1.0 621 107 65.9 32.29 541.4 6.9 <1.0 10.1 108 59.06 61.43 1165 <1.0 <1.0 119.3 109 17.97 60.7 3260 12.3 4.2 226.8 110 138.9 40.75 9459 2.2 1.1 — 111 284.8 240.8 1135 >30.0 19.8 — 112 178.9 76.78 3581 5 5 — 113 75.94 29.93 1370 13.6 <1.0 — 114 13.57 6.953 431.3 23.3 <1.0 — 115 6.213 8.144 229.4 9 <1.0 <1.0 116 65.02 22.02 454.6 <1.0 <1.0 383.1 117 Inactive 14410 4734 >30.0 >80.0 — 118 29.61 6.603 269.1 >30.0 <1.0 — 119 32.39 7.547 75.39 18.4 <1.0 — 120 35.85 7.498 224.6 >30.0 <1.0 — 121 443.1 221 522.6 5.4 — 242.7 122 370.9 261.2 1303 5.4 — 822.3 123 442.4 145.2 857 7.3 — — 124 349 144.2 1357 19.7 — 20.4 125 39.76 8.083 506.1 — — — 126 118.2 24.04 372.4 1.7 <1.0 1238.7 127 169.1 105.3 338.7 12.2 7.3 9.1 128 31.45 6.054 310.6 12.7 1.5 735.4 129 680 1445 856.7 >30.0 20.3 5.4 130 72.2 20.17 202.1 >30.0 9.2 467.2 131 17360 17480 1557 >30.0 >75.0 1.9 132 344.1 63.1 736.2 <1.0 <1.0 2316 133 10120 3997 inactive >30.0 <1.0 <5.4 134 255.7 72.67 2007 10.5 <1.0 142.1 135 100000 77800 inactive >30.0 >80.0 3 136 732.7 94.52 3591 7.8 8.6 1292.9 137 1004 187.9 1518 1.6 <1.0 — 138 276.8 173.2 1278 4.8 <1.0 890.6 139 152.9 20.63 405.1 4.3 <1.0 238.4 140 272.2 59.24 6373 9.6 >77.0 1393.8 141 260.1 47.47 8043 4.2 66.5 1500.6 142 50000 13770 46170 1.4 <1.0 ND 143 242.1 63.12 671.6 1.6 <1.0 1421.5 144 1761 367.7 2083 4.1 <1.0 1470.8 145 266.3 60.65 1214 4.9 <1.0 362.8 146 1814 535.8 5732 9.6 63 98.3 147 18300 4809 21220 3.1 >90.0 108.6 148 4575 601.1 18170 >30.0 21.8 — 149 839.5 408.6 6235 >30.0 8.8 — 150 6229 3824 7873 2.4 3.9 — 151 52.67 19.05 3720 3.5 <1.0 — 152 4038 2205 796.2 3.3 <1.0 — 153 52.91 126.8 18360 9.4 <1.0 57.9 154 101.7 159.9 765.7 8 <1.0 <1.0 155 3113 1235 27970 10.7 <1.0 <1.0 156 1576 686.7 67630 5.6 <1.0 <1.0 157 7430 3350 50000 6.8 <1.0 <1.0 158 3556 1555 7899 18.8 <1.0 <2.4 159 320.5 91.59 3523 — — — 160 270.1 96.06 9278 >30.0 >79.0 <1.0 161 1536 512.5 47050 >30.0 >82.0 5.4 162 894.7 368 32080 >30.0 >83.0 <1.0 163 246.5 93.88 6497 >30.0 >72.0 231.9 164 809 594.4 40420 >30.0 >79.0 238.3 165 4091 933.7 50000 >30.0 >80.0 1.8 166 179.4 72.73 8762 >30.0 >70.0 426.6 167 469 223.2 22690 >30.0 >76.0 1.5 168 224.3 85.97 1194 >30.0 17.3 1.4 169 629.7 301 631.5 — 2.8 204.2 170 168.1 122.2 3773 13.3 2.7 591 171 240.4 47.51 154.3 2.8 >72.0 31.3 172 202.8 121 967.8 >30.0 2.3 12.3 173 2526 — 100000 — >77.00 365.72 174 265 132.7 7636 >30.00 >68.00 49.89 175 1618 731 11780 >30.00 66.62 — 176 282.7 90.23 556.2 — <0.10 ND 177 902.1 — 9038 — >74.00 — 178 918 444.9 5312 — — — 179 1696 451.1 100000 19.4 <1 <1 180 1821 931.4 2454 >30 43.7 3.8 181 1923 1584 830.4 >30 17.3 36.3 182 1479 459 1439 10.8 1.1 0 183 131.1 49.84 794.5 >30 28.4 <1 184 820.6 171.3 18050 >30 56.1 292.5 185 1144 352.5 50000 15.1 3.4 35.5 186 572.8 207.2 12440 — — — 187 4588 1605 724.6 0 0 0 188 601.5 — 7201 21.5 <1 1247 189 357.9 — 6514 >30 >78 212.5 190 248.5 — 10680 >30 >77 0 191 172.3 — 4337 26.7 2.4 606.4 192 174.4 11990 24.7 <1 0 193 813.6 — 8290 >30 >81 161.6 194 701.5 — 100000 6.9 <1 1078.4 195 275.8 — 781.4 8.8 <1 148 196 100000 — 100000 18 <1 85.9 197 65140 — 2384 >30 >77 3.5 198 263.6 13840 >30 >75 6.1 199 108.8 16510 5.6 1.4 1442.8 200 60.03 15370 0 7.2 1070.1 201 1619 17280 12.6 <1 13.8 202 120.4 3267 >30 57.8 415.1 203 17.23 512.8 29.3 1.2 749.5 204 14.26 257.1 >30 <1 <1 205 177.2 1061 0 2.2 455.6 206 48.9 19640 7.1 44.1 427.6 207 145.4 168.8 >30 58.1 <1 208 65.92 21750 13.2 35.5 247.9 209 537.7 2492 4 <1 <1 210 14470 100000 2.7 <1 0 211 3878 43090 >30 >83 227 212 3888 15800 >30 >79 188.7 213 130.2 1299 18.9 <1 <1 214 18.82 339.4 21.2 <1 333.1 215 177.5 763.8 6.3 <1 492.1 216 42.23 832.1 6.7 <1 1.1 217 531.3 17830 >30 >83 130 218 1684 16250 >30 >69 100.4 219 370.7 3288 >30 6.3 3.3 220 156.1 4247 12.6 9.4 127.4 221 45.47 1504 22.2 <1 133.1 222 22.83 820.6 27.2 7.1 757.6 223 58.02 7119 — 3.4 1178.5 224 46000 100000 0 >74 <1 225 19.75 916.9 23.3 <1 735.2 226 208.1 8446 — — — 227 66.45 16000 — — — 228 194.9 11220 — — — 229 232.4 19250 — — — 230 98.17 50000 — — — 231 22.74 850.5 18 <1 364.4 232 53.43 1065 >30 9.2 7.6 233 162.4 6306 >30 >76 155.1 234 2219 18920 1.4 <1 874.2 235 24450 100000 3.4 <1 563.9 236 1198 50000 >30 >78 <1 237 39510 100000 >30 >65 3.3 238 56.84 214.8 >30 <1 438.9 239 17.47 242.2 >30 <1 79.9 240 52.88 443.6 >30 31.6 <1 241 334.7 6061 >30 >74 18.6 242 488.7 373 >30 >74 <1 243 676.9 14600 >30 >76 382.2 244 1635 7873 >30 >76 216.2 245 3817 10410 >30 >61 11.7 246 1257 645.9 >30 >72 1.5 247 81.83 1903 16.7 <1 846 248 329.2 23380 10.6 51.2 ND 249 575.5 14760 7.9 >82 1066.3 250 549 2911 >30 9.5 14.3 251 2374 100000 1.3 <1 <1 252 100000 100000 — — — 253 257.8 8463 >30 >80 352.9 254 206.2 3012 >30 53.1 5.7 255 181.9 12950 2.3 >88 1044.2 256 500.7 8776 3.5 <1 663.6 257 60.78 2719 >30 >84 1.5 258 34.3 2541 24.4 0 0 259 24.18 655.9 2.6 0 0 260 62.41 5024 2.1 0 0 261 60.03 1871 1.7 0 0 262 213.8 5399 4.3 0 0 263 58.82 1512 >30 0 0 264 56.56 1321 4.5 0 0 265 132 2064 1.9 0 0 266 58.28 3013 2.8 0 0 267 58.15 3878 8 0 0 268 211.1 4985 2 0 0 269 389.9 3874 >30 0 0 270 348.4 3254 >30 0 0 271 612.6 16140 14 0 0 272 52.07 5227 21.9 11.8 625.8 273 18.9 4164 >30 <1 0 274 40.61 4900 21.4 63.8 1007.1 275 44.22 4039 19.9 65.6 426.8 276 222.2 21760 5.8 34.8 973.2 277 82.48 3631 3.8 19.9 354 278 110.2 8000 2.6 >83 1383.7 279 50.5 1317 15.8 >84 49.1 280 152.4 1609 >30 73.6 49.1 281 27.38 2180 3 >86 33.4 282 71.4 4536 4.1 >86 598.2 283 69.1 15090 2.7 70.7 1184.7 284 182.9 9354 7.5 36.6 280.6 285 59.71 1506 >30 41.6 159.4 286 53.68 11770 2.4 35.6 863.4 287 58.44 2083 4.3 <1 21.7 288 283.9 1258 1.2 <1 18.8 289 86.73 9216 1.7 <1 141.1 290 64.38 4806 3 63.1 310.7 291 210.9 4975 3.1 0 772.9 292 1137 15560 1.7 0 3.4 293 146.3 6103 22.9 0 241.2 294 2046 4586 8.4 0 0 295 162.2 23740 10.6 0 638.9 296 477.2 18600 7.1 0 509.8 297 150.1 11020 1.6 0 936.2 298 949.5 50000 1.5 0 148.2 299 55.34 5830 — — — 300 32.74 3792 — — — 301 205.5 50000 >30 <1 0 302 16.37 1595 12 <1 0 303 6.446 237.6 >30 0.7 0 304 19.2 853.5 — — — 305 483.3 4727 — — — 306 100.2 8864 — — — 307 190.9 39900 — — — 308 23.56 2099 — — — 309 553.9 3824 — — — 310 413.7 3373 — — — 311 53.26 198.2 — — — 312 1853 500000 — — — 313 39.03 1429 — — — 314 429.4 780 — — — 315 137 100000 — — — 316 412.1 8154 — — — 317 201.7 8572 — — — 318 — — — — — — 319 — — — — — — 320 — — — — — — 321 — — — — — — 322 — — — — — — 323 — — — — — — 324 — — — — — — 325 — — — — — — 326 — — — — — — 327 — — — — — — 328 — — — — — — 329 — — — — — — 330 — — — — — — 331 — — — — — — 332 1519 859 494.9 >30.0 >64.0 31.9 333 411.2 238.7 213.5 — — — 334 19.73 8.213 88.72 >30.0 30 14.9 335 17.74 14.44 28.21 >30.0 2.8 <2.2 336 40.99 10.17 19.71 >30.0 <1.0 <3.6 337 6232 1574 3229 >30.0 >68.0 — 338 65.86 N/A 150.5 — 0.32 140.79

TABLE 4 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 339 9.19 0 299.9 >30.0 <1.0 <1.0 340 616.5 266.1 15200 >30.0 >68.0 5.2 341 82.95 69.08 620.5 12.4 <1.0 ND 342 349.8 129.9 6710 >30.00 31.01 12.72 343 — 20.85 447.6 >30 13.4 <2.5 344 1505 0 3539 19.2 <1.0 238.8 345 3832 2.342 7622 <1.0 — — 346 48.38 18.57 574.1 — — — 347 1719 667.7 14370 >30.0 >70.0 317.6 348 2446 1458 23930 >30.0 >72.0 289.4 349 26780 14220 16230 >30.0 >68.0 2.5 350 1003 546.2 10110 — >68.0 — 351 2162 155.9 433 — — — 352 100000 18750 11480 — >72.0 343.6 353 2514 588.7 50000 >30.0 0 NOT_DETERMINED 354 1907 687.4 15790 >30.0 >74.0 292.3 355 28750 2022 2516 >30.0 >77.0 — 356 65.49 24.98 1044 >30.00 <0.10 NOT_DETERMINED 357 2197 862.9 8193 >30.00 >70.00 40.53 358 2184 878.5 6888 — >72.00 1209.2 359 129.7 23.44 1328 14.9 0.06 — 360 736 170.2 1696 19.2 0.73 635.05 361 6441 1575 100000 >30.00 <0.10 199.84 362 81.71 8.86 1278 7.4 0.6 98.56 363 860.2 449.6 4610 — 57.74 8.76 364 100000 4582 50000 >30 2.7 75.9 365 30610 481.5 126.9 0 55.9 <1.8 366 3759 1235 12120 0 14.8 47.8 367 104.7 29 326.5 12.8 0 ND 368 2195 1283 3401 >30 6.5 2.9 369 2942 1228 17820 26 15.7 55 370 4115 296.2 1456 >30 27.3 <1 371 2473 471.6 31140 >30 45.3 1.2 372 1049 540.2 50000 >30 <1 <1 373 45.72 349.6 1735 >30 3.3 <6.3 374 12930 4183 50000 >30 52.5 <1.6 375 714.8 389.1 50000 >30 13.1 <1.1 376 198.6 115 460.1 0 0 0 377 384.1 3.841 1177 0 0 0 378 1546 — 17200 >30 >70 2.3 379 — 2956 7400 23 <1 31 380 — 310.5 1365 >30 2.6 <10.7 381 — 263.8 652.7 >30 44.9 4 381 — 8.438 2938 — — — 383 — 62.31 1837 — — — 384 — 1322 100000 — — — 385 — — — — — — 386 — — — — — — 387 1838 315.8 262.6 >30.0 1.4 <4.1 388 88.15 28.19 120 — 11.8 2 389 65.27 21.83 90.23 — <1.0 <9.7 390 115.8 75.16 60.97 >30.0 4.2 25 391 56.28 12.26 126.8 >30.00 10 ND 392 46.08 22.04 122.6 >30.00 33.81 133.39 393 30.71 9.305 19.62 — 6.26 <1.10 394 438.5 129.8 107.5 >30 <1 <1 395 46000 — 53.39 >30 11.3 3.9

TABLE 5 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 396 — 144.1 15140 >30 46.4 207.8 397 232.3 267.8 Inactive >30.0 <1.0 — 398 194.1 104.9 2471 >30.0 <1.0 — 399 522.2 219.3 100000 >30.0 <1.0 ND 400 294 165.2 100000 >30.0 <1.0 1464.6 401 190.2 86.7 1271 >30.0 <1.0 296.7 402 343.1 139.9 53210 >30.0 1.9 222.8 403 262.2 125.3 100000 >30.0 3.1 ND 404 1644 681.4 50000 >30.0 0.9 19.5 405 665.7 298.4 100000 2.4 <0.1 ND 406 13.07 8.463 216.5 19.4 <1 ND 407 104.2 52.41 50000 >30 <1 0 408 304.7 10.04 100000 0 0 0 409 105.7 52.89 50000 0 0 0 410 36.3 2.488 100000 0 0 0 411 108.7 61.4 100000 0 0 0 412 39.88 6.574 100000 0 0 0 413 53.99 29.35 2730 0 0 0 414 38.71 14.21 584.2 — — — 415 18.63 21.43 15870 — — — 416 21.64 39.63 930.6 — — — 417 36.45 — 319 0 <1 <1 418 — 7.934 700.2 — — — 419 — 82.94 30380 — — — 420 — 144.2 30220 — — — 421 — 18.91 18940 — — — 422 — 5.866 1870 — — — 423 — 50.42 15400 — — — 424 — 22.99 12510 — — — 425 — 48.43 5000 — — — 426 — 29.43 6418 — — — 427 — 75.16 30600 — — — 428 — 27.77 11000 — — — 429 — 10.52 12570 — — — 430 — 58.45 4622 — — — 431 — — — — — — 432 — — — — — — 433 — 17.07 6100 — — — 434 Inactive 45000 >46000 uM 11.3 <1.0 ND 435 3657 364.6 >30 30.8 11 436 1966 891.9 12450 >30.0 52.9 767.2 437 44.34 14.4 529.3 >30.0 <1.0 3.1 438 169.3 174.1 100000 27.2 <1.0 <1.0 439 843.9 37.93 714.1 >30.0 1.4 18.4 440 2179 786.6 100000 >30.0 <1.0 ND 441 685.5 319.1 516.2 >30.0 <1.0 <1.0 442 1992 449.1 100000 — — — 443 68.27 44.55 302.4 >30.0 3.3 644.7 444 279.7 106.1 427.8 12.8 <1.0 406.6 445 734.7 269.4 630.3 — — — 446 — 1669 6805 >30 1 <16.2 447 — 139.3 16340 26.7 7.4 1377.3 448 50000 20020 100000 >30.0 0.4 ND 449 100000 6346 100000 2.9 0.2 ND 450 312.8 88.3 1108 17.9 <0.10 — 451 50000 8408 100000 — <0.10 ND 452 2917 1186 100000 >30 <0.1 ND 453 100000 3996 479.6 14.9 29.3 <0.5 454 1627 301.2 100000 0 <0.1 ND 455 2060 1013 6581 — — — 456 3156 931.4 1313 397.4 3.9 <0.9 457 2543 667.9 50000 >30 <1 <13.5 458 456.7 22.05 42820 — — — 459 443 88.47 100000 — — — 460 100000 147.7 50000 9.8 <1 927.1 461 — 1766 100000 9.9 43.8 ND 462 — 569.1 100000 8.2 <1 2112.3 463 — 3417 100000 17.1 <1 306.8 464 137.2 45.3 10680 18.5 <1 1021.6 465 525.5 113.4 50000 >30 <1 953.8 466 18.31 3.626 7762 >30 6.6 174.1 467 23.69 15.04 1776 17.6 <1 792 468 114 60.19 7448 0 0 0 469 125.5 52.96 100000 18.6 42.7 814.3 470 217.2 163.6 20420 0 0 0 471 156.3 60.34 6030 0 0 0 472 71.56 20.6 4764 0 0 0 473 39.3 16.87 3115 0 0 0 474 24.99 17.6 3382 0 0 0 475 24.5 11.1 459.7 0 0 0 476 473.8 231.3 19430 — — — 477 2371 1615 4288 0 0 0 478 292.5 — 49380 >30 >66 0 479 141.1 — 1250 11.2 <1 <13.3 480 8715 — 3258 11.9 <1 0 481 851.2 — 3462 10 35.9 <1 482 2749 — 6901 0 39.7 35.8 484 306 — 1312 0 <1 81.6 485 81.01 — 562.6 0 <1 13 486 77.87 — 299.5 0 <1 <4.6 487 — 469.4 100000 30.5 37.9 804.3 488 — 4845 100000 >30 <1 ND 489 — 55.86 100000 >30 <1 0 490 — 2984 100000 >30 <1 ND 491 — 51.4 22990 4.7 53 1585.7 492 — 944.1 2384 >30 <1 <1 493 — 805.4 100000 >30 <1 ND 494 — 433.3 100000 >30 <1 ND 495 — 2972 100000 >30 <1 192.2 496 — 130.1 2358 >30 <1 <5.3 497 — 132.1 6485 12.9 <1 0 498 — 3736 100000 30.4 57.5 1434.3 500 — 1499 100000 — 48.1 1393.2 501 — 259.7 100000 7.4 53.6 993.7 502 — 18.53 682.2 7.2 <1 ND 503 — 542.7 47880 23.3 <1 ND 504 — 187.8 500000 >30 27.4 960.1 505 — 16.78 12320 >30 <1 0 506 — 27.17 4579 >30 1.1 <1 507 — 221.8 8798 >30 56.7 720.1 508 — 75.14 3176 >30 58.4 1010.7 509 — 100.3 43950 >30 >68 1289 510 — 25.33 100000 8.5 <1 1361 511 — 8.5 100000 — — — 512 — 19.75 18500 — — — 513 — 26.5 39070 22.5 <1 0 514 — 147.4 100000 5.2 <1 ND 515 — 7.082 50000 12.1 0 1471.9 516 — 465.9 100000 7 0 1285.5 517 — 12.33 100000 — — — 518 — 2819 1876 — — — 519 — 5.728 722 — — — 520 — 38.03 21200 — — — 521 — 117.8 4237 — — — 522 — 260.1 100000 — — — 523 — 1033 9590 — — — 524 — 40.01 6470 — — — 525 — 191.5 29140 — — — 526 — — — — — — 527 — — — — — — 528 — — — — — — 529 — — — — — — 530 — — — — — — 531 — — — — — — 532 — — — — — — 533 — — — — — — 534 — — — — — — 535 — — — — — — 536 — — — — — — 537 — — — — — — 538 — — — — — — 539 — — — — — — 540 — — — — — — 541 — — — — — — 542 — — — — — — 543 — — — — — — 544 482.5 385.6 174.1 >30.0 <1.0 <17.5 545 118.1 98.4 499.1 >30.0 <1.0 20 546 63.1 37.07 28.44 — <1.0 <6.2 547 4.101 2.69 46.71 >30.0 <1.0 — 548 27.94 6.836 490.2 >30.0 43.5 3.3 549 7.232 3.995 541.4 >30.0 43.8 199.1 550 310.3 140.9 5548 ND 2.5 145.9 551 31.9 21.51 111 >30.0 1.6 12.7 552 1111 97.77 2908 >30.0 2.3 4.6 553 92.85 77.96 553.9 >30.0 1.3 <1.0 554 707.9 298.8 12330 >30.0 8.6 543.6 555 706.7 301 50000 — — — 556 21400 6332 50000 — 20.7 100.7 557 58.16 13.61 420.4 25.9 1.4 529.2 558 372.3 101.7 1264 20.8 3.4 0 559 655 386.8 289.6 5.3 5.3 275 560 1837 377.1 84.97 >30 11.5 0 561 22.07 9.823 44.49 >30.00 1.51 377.44 562 50000 1285 100000 >30.00 <0.10 <30.20 563 3562 897.2 50000 0 11.3 0 564 18.29 5.258 985.5 0 0 0 626 — 23.33 26260 >30 47.9 78.6 627 — 17.14 3535 >30 29.9 80.3 628 — 97.97 557.9 14 <1 876.8 629 — 163.6 22780 >30 <1 983.9 630 — 57.2 28400 >30 <1 810.6 631 — 45.16 100000 9.1 <1 2.5 632 — 12.96 9384 7.6 <1 1241.2 633 — 48.49 50000 12.1 <1 382 634 — 1221 220000 2.8 11 339.4 635 — 19.16 26730 — — —

TABLE 6 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 565 1332 445.7 19430 >30.0 <1.0 ND 566 816.3 526 15670 >30.0 <1.0 ND 567 3737 7805 2906 >30 56.7 0 568 14570 4370 50000 — — — 569 100000 14680 6560 — — — 570 28740 — 2694 >30 3.7 1.7 571 228.3 68.11 170.8 >30.0 4 27.8 572 557.8 67.75 25.37 — — —

TABLE 7 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 573 237.2 54.66 794.5 >30.0 <1.0 >1625.0 574 536.5 115.5 1477 25.8 <1.0 222.9 575 337.5 218.1 1059 22.7 3.3 3 576 — 153.7 8427 — — — 577 — 197.2 100000 — — — 578 2769 2174 18080 18.5 <1.0 <4.8 579 719.3 418.2 28100 — — — 580 1118 1231 4304 — 5.4 654.6 581 1060 632.2 1824 — 5.2 ND 582 1850 992 10020 5.5 <1.0 ND 583 37450 Inactive 3575000 12.2 <1.0 ND 584 2619 494.9 1452 17.8 <1.0 31.6 585 7647 1092 171.4 19 <1.0 79.3 586 8025 4802 10510 >30.0 >77.0 220.3 587 688.6 245 6702 18.2 <1.0 ND 588 5312 1424 100000 18.5 <1.0 83.1 589 100000 913.8 100000 — — — 590 1083 217.8 4761 5.3 2 1345.4 591 1909 1066 2095 6.2 <1.0 1336.2 592 50140 — 37960 >30 12.7 <1 593 100000 — 100000 — — — 594 — 4275 100000 19.6 <1 144.8 595 — 683.3 100000 — — — 596 — 349.9 26820 — — — 597 — 490.1 100000 — — — 598 — 5124 100000 — — — 599 — 43840 6384 — — — 600 — 251.4 14470 — — — 601 — 431.3 7223 — — — 602 3348 302.4 127.4 >30.0 <1.0 <3.1 603 107.5 82.18 195.4 >30.0 1.1 <6.4 604 374 107.4 33.56 6.5 <1.0 <12.3 605 159.4 60.87 111.3 15.6 <1.0 231.6 606 120 50.73 79.18 >30.0 1.9 70.9 607 5635 113.9 116.2 >30.0 4.9 <1.9 608 106 34.84 210 >30.0 7 12.2 609 28990 351.9 335 >30.0 4.5 ND 610 150.4 50.9 257 >30 3.9 0 611 2232 225.6 234 >30.0 1.6 <3.6

TABLE 8 Game- Microsomal tocyte Asexual Stability Sol @ PAMPA EC50 EC50 mTOR (rat) T_(1/2) pH 7.4 Permeability # (nM) (nM) (nM) (min) (ug/mL) (×10⁶ cm/s) 612 16870 5660 11410 2.2 54.4 — 613 100000 14080 13650 >30.0 37.7 2.9 614 11290 3534 12050 >30.0 58.6 — 615 100000 3800 29840 >30.0 40 3.1

Example 7

This example demonstrates a synthesis of compounds, in accordance with an embodiment of the invention.

N-methyl isopropyl carbamate 4 of 6-bromo-4-chloroquinolin-3-amines 2, one example of the key intermediate, can be synthesized from commercially available 6-bromo-4-chloroquinolin-3-amine 1 in three steps: stannous chloride mediated reduction of 1 followed by carbamate synthesis through isopropyl chloroformate gave 3, subsequent methylation provided 4, as shown in Scheme 6.

One aspect of the invention provides a process of making a compound of structure 6 in a high boiling point organic reaction medium under microwave irradiation, further coupling the compound of structure 6 with the boronic species to obtain the compound of structure 7, as shown in Scheme 7.

Preparation of 6-bromo-4-chloroquinolin-3-amine 2

To a mixture of 25 g (85 mmol) of 6-bromo-4-chloro-3-nitroquinoline 1 with 500 mL of EtOH and 75 mL of AcOH was added 98 g (435 mmol) SnCl₂ dihydrate as one portion. The reaction mixture was then refluxed for 3 hours. The reaction mixture was cooled to 0° C. and the pH was adjusted to basic with saturated NaHCO₃. The mixture was extracted with 1 L of EtOAc three times. The EtOAc layers were combined, dried over MgSO4 and concentrated. The resulting solid was triturated with Et₂O to provide 14.3 g of 2 as a pale-yellow solid.

Preparation of isopropyl (6-bromo-4-chloroquinolin-3-yl)carbamate 3

Under an inert atmosphere, to a stirred solution of 2 (7.98 g, 31 mmol) in anhydrous DCM (140 mL) at 0° C. was added pyridine (37.2 mmol), followed by 34.1 mL isopropyl chloroformate (34.1 mmol, as 1M solution in PhMe) dropwise. The solution was stirred at 0° C. for a further 15 min before the ice bath was removed and stirring continued at rt for 4 hours. Then 200 mL DCM was added at 0° C., followed by an equal volume of saturated NaHCO₃ and several drops of MeOH. The mixture was extracted with 200 mL of DCM three times. The organic layers were combined, dried (MgSO₄) and concentrated. The resulting orange red mixture was triturated with Et₂O (100 mL×3), providing 7.0 g pure product 3 as a pale-orange solid.

Preparation of isopropyl (6-bromo-4-chloroquinolin-3-yl)(methyl)carbamate 4

Under an inert atmosphere, to a stirred solution of 3 (7 g, 20.3 mmol) in anhydrous THF (100 mL) at 0° C., was added portion-wise NaH (1.63 g, 40.7 mmol, 60% dispersion in mineral oil). After 15 min, iodomethane (3.8 mL, 61.1 mmol) was added dropwise and the resulting mixture was stirred at 0° C. for 60 min and then at rt for 18 hours. At 0° C., 200 mL saturated NH₄Cl was added. The mixture was extracted with 200 mL of EtOAc three times. The EtOAc layers were combined, dried (MgSO4) and concentrated. The residual mixture was then passed through a silica gel column, and the product eluted with DCM and MeOH. The resulting pale orange oil was left under vacuum, providing 6.7 g product 4 as pale-yellow solid.

Preparation of Compound 7 Through One-Pot Two Step Synthesis

In a 0.5 mL-2.0 mL conical shaped microwave reaction vial, isopropyl (6-bromo-4-chloroquinolin-3-yl)(methyl)carbamate 4 (36 mg, 0.1 mmol) and 1.0 mL of 3-pentanol were mixed, followed by aniline 5 (0.2 mmol) and 75 uL HCl (0.3 mmol, 4.0 M in dioxane). The mixture was microwave irradiated at 230° C. for several minutes. 0.5 mL DMF and 0.5 mL 1M K₃PO₄ (106 mg K₃PO₄ in 0.5 mL H₂O)^(a) were added. With stirring, the air was displaced with nitrogen. Boronic acid/ester (0.2 mmol) and Pd(PPh₃)₄(0.003 mmol) were introduced. The mixture was microwave heated at 150° C. for 10 minutes. The reaction mixture was cooled and 5 mL of saturated NaHCO₃ was introduced. The reaction mixture was extracted with three 7 mL portions of DCM. The organic extract was dried (MgSO₄), filtered and concentrated. To obtain a pure product, the residual mixture was passed through a silica gel column, and the product 7 was eluted with DCM and MeOH.

^(a)Note: For difficult examples, molar ratio (4:5:HCl)=1:4:5, volume of 1M K₃PO₄=0.7 mL.

Preparation of Compound 6 Through Conventional Reflux

In a round bottomed flask, N-methyl carbamate 4 (3 mmol) and 10 mL cyclohexanol were mixed, followed by aniline 5 (9 mmol) and HCl (12 mmol, 4.0 M in dioxane). The mixture was refluxed for 4 hours (or until reaction was complete). Then at 0° C., 30 mL of saturated NaHCO₃ was introduced. The mixture was then extracted with three 30 mL portions of DCM. The organic extract was dried (MgSO₄), filtered and concentrated. The resulting mixture was passed through a silica gel column, and product 6 was eluted with DCM and MeOH.

All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context. 

The invention claimed is:
 1. A compound of formula (I):

wherein A is CH, B is CR³═CR⁴, R¹ is selected from 4-methylsulfonylphenyl, 4-ethylsulfonylphenyl, 2-methyl-4-methylsulfonylphenyl, and 3-piperazinylmethyl-4-methylsulfonylphenyl, R² is selected from 4-chlorophenyl, 4-fluorophenyl, 3-cyano-4-chlorophenyl, 3-hydroxy-4-chlorophenyl, 3-amino-4-chlorophenyl, 4-aminomethylphenyl, and 3-methoxy-4-chlorophenyl, R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl, and R⁵ is C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, or hydroxyl, or a pharmaceutically acceptable salt thereof.
 2. The compound or salt of claim 1, wherein the compound is selected from:


3. A compound of formula (I):

wherein A is CH, B is CR³═CR⁴, R² is selected from 4-chlorophenyl, 4-fluorophenyl, 4-dimethylaminomethylphenyl, 3-methylphenyl, 3-methoxyphenyl, 3-cyanophenyl, phenyl, 5-indolinone, 5-isoindolinone, 3-cyano-4-chlorophenyl, 4-cyanophenyl, 4-(2-dimethylaminoethyl)phenyl, 4-azetidinylphenyl, and 4-ethylphenyl, 2-amino-5-pyridyl, and R¹ is selected from 4-morpholinocarbonylphenyl, 4-dimethylaminocarbonylphenyl, 4-(3,5-dimethylaminomorpholino)carbonylphenyl, 4-(4-methylpiperazinyl)carbonylphenyl, 4-piperazinylcarbonylphenyl, 4-piperidinylcarbonylphenyl, 4-cyclopentylaminocarbonylphenyl, 4-azetidinylcarbonylphenyl, 4-(4-hydroxyethylpiperazinyl)carbonylphenyl, 4-(N-methyl-N-cyclopropylamino)carbonylphenyl, 4-cyclobutylaminocarbonylphenyl, 2-trifluoroethylaminocarbonylphenyl, 4-(2-dimethylaminoethylaminocarbonyl)phenyl, 4-(4-[2-dimethylaminoethyl]piperazin-1-yl-carbonyl)phenyl, 4-cyclopropylaminocarbonylphenyl, 4-(1-[2-dimethylaminoethyl]piperidin-4-amino)carbonylphenyl, 4-(N-(1-(2-hydroxyethyl)azetidin-3-ylamino)carbonylphenyl, and 4-(pyrrolidinyl-3-amino)carbonylphenyl, R³ and R⁴ are independently selected from hydrogen, hydroxyl, OR⁵, halogen, optionally substituted C₆₋₁₀ aryl, and optionally substituted C₁₋₆ alkyl, and R⁵ is C₁₋₁₂ alkyl, C₆₋₁₀ aryl, halogen, or hydroxyl.
 4. The compound or salt of claim 3, wherein the compound is selected from:


5. A pharmaceutical composition comprising a compound or salt of claim 1 and a pharmaceutically acceptable carrier.
 6. A method of blocking transmission of a Plasmodium parasite or treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a first compound of claim 1, optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat.
 7. A pharmaceutical composition comprising a compound or salt of claim 3 and a pharmaceutically acceptable carrier.
 8. A method of blocking transmission of a Plasmodium parasite or treating malaria by killing or arresting the growth of Plasmodium organisms in a mammal, wherein the Plasmodium organisms are in a gametocyte stage comprising administering to a mammal in need of such treatment, a therapeutically effective amount of a first compound of claim 3, optionally in combination with an antimalarial compound selected from elesclomol, NSC174938, NVP-AUY922, maduramicin, narasin, alvespimycin, omacetaxine, thiram, zinc pyrithione, phanquinone, bortezomib, salinomycin sodium, monensin sodium, dipyrithione, dicyclopentamethylene-thiuram disulfide, YM155, withaferin A, adriamycin, romidepsin, AZD-1152-HQPA, CAY10581, plicamycin, CUDC-101, auranofin, trametinib, GSK-458, afatinib, and panobinostat. 